Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ; test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.
The present study was conducted to investigate the effects of fusion and/or activation protocol on in vitro development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine fetal fibroblast cells were transferred into the perivitelline space of enucleated in vitro matured oocytes. Cell fusion and activation were induced simultaneous fusion/activation (SA) or delayed activation (DA) with or without cytochalasin B (CB) treatment with electic pulses in 0.28 M mannitol-based medium. The SCNT embryos were cultured in vitro for 7 days and stained with Hoechst 33342 to determine the number of nuclei. After 7 days culture, cleavage and blastocyst formation rates were 72.4% and 7.6% in SCNT and 76.3% and 20.4% in parthenotes. To examine the effect of electric field strengths on development of SCNT embryos, oocytes were fused two pulses of 110 V/mm, 130 V/mm or 150 V/mm for 30 sec post-injection. The fusion and cleavage rates in 130 V/mm group (70.2% and 72.6%) and 150 V/mm group (72.6% and 70.5%) were higher (P<0.05) than 110 V/mm group (47.1% and 48.6%), respectively. However, the rate of embryos developing to the blastocyst stage (8.1%, 9.7% and 10.7%) were not different among three groups. The cleavage rates and the blastcyst formation rates were not different among three treatment groups (SA group, 71.4% and 9.7%; SA+CB treatment group, 74.7% and 8.0%; DA+CB treatment group, 70.8% and 11.2%, respectively). And, no different in the number of cells in blastocysts was observed among the three groups (22.5$\pm$12.8, 23.3$\pm$11.2 and 21.6$\pm$10.4, respectively). These result suggest that two pulses of 130 V/mm or 150 V/mm for 30 sec with SA treatment or DA treatment are enough for fusion/activation of porcine somatic cell nuclear transfer (SCNT) embryos to develop to the blastocyst stage.
This study was conducted in order to grasp the condition of the student's knowledge and attitude about tuberculosis. And to offer the basic materials for the prevent tuberculosis in elementary school. The objects were comprised 395 of volunteered 6th grade students who study in two elementary schools. The material of this study was the questionaire(chronbach $\alpha=.9016$) suited to the purposed of this research which has been made through studying references. All the questionaire were collected immediately without explanation. The data was collected from 13rd to 25th of May on 1994. Analysis of the data was done utilizing SPSS for percentage, mean, ANOVA and pearson-Correlation Coefficient. The Result are as follows; 1. General features of the objects of study. Sex distribution was similar. Salary of 1000-1500 thousand won were highest (40.2%), nuclear family was highest (87.1%), fathers of graduated high school were the highest(44.0%), mothers of graduated high school were the highest(56.8%). And there was nobody about tuberculosis patient in one's close relatives were highest(95.4%). 2. The conditions of attitude about tuberculosis. Having experience of tuberculin test were highest(59.0%). Less than 9mm indulation among the tuberculin tested group were 85.1%, more than indulation were 14.9%. Have a B. C. G. injection among the less than 9mm indulation were 83.2%. Leave alone among the more than 10mm indulation were 56.0%. Think it too much trouble to do not tuberculine test were 63.0%. Do not take a chest X-ray in the past were 60.3%. And take a chest X-ray in the past were 39.7%. Health educated group was 74.3%. If tuberculosis sign was developed, notified to parent was 73.8%. 3. The condition of knowledge about tuberculosis. When 5 points was given to 'very affiming' and 1 point was given to 'very deny', the total average was 3.54. And symptom of tuberculosis secion was 3.67, vaccination of tuberculosis section was 3.66, tuberculine test section was 3.56, epidemiology of tuberculosis section was 3.54, infection of tuberculosis section was 3.38. And every section showed affirmative correlation(P<.001). 4. Correlation between the general features and attitude variables. High incomed group may have more attitude on tuberculin test than low incomed group($x^2=16.$ 190, P<.01). High educated group may have more attitude on tuberculin test than low educated group(Father : $x^2=28.530$, P<.001, Mother: $x^2=26.060$, P<.001). High educated group may have more attitude on health education than low educated group(Father: $x^2=20.$ 767, P<.000, Mother: $x^2=10.639$, P<.05). Nuclear family may have more attitude on notify to parent than others($x^2=51.45$, P<.000). Tuberculosis patient in one's close associates have more attitude on notify to parent than others($x^2=51.$ 45, P<.000). 5. Difference between the general features and knowledge of tuberculosis. High incomed group were highest score in knowledge (F=3.99, P<.01). High educated group were highest score in knowledge(Father : F=8.81, P<.000, Mother: F=9.09, P<.000). 6. Difference between the attitude and knowledge of tuberculosis. Tuberculin tested group were highest score in knowledge(t=9.88, P<.000). Taken chest X-ray group were highest score in knowledge (t=2.07, P<.05). Received health education group were highest score in knowledge(t=6.83, P<.000). Notified symptoms to teachers and parent group were highest score in knowledge(F=3.89, P<.01).
Purpose : In order to evaluate the hypoxia-ischemia(H-I) induced neurotoxicity and the protective effect of xanthine oxidase(XO) inhibitor(allopurinol), cell number, cell viability, lactate dehydrogenase(LDH), protein synthesis(PS) and protein kinase C(PKC) activity were measured in cerebral neurons and astrocytes. Methods : Cytotoxic effect was measured by in vitro assay at 12-72 hours after H-I on cerebral neurons and astrocytes derived from 7-day old neonatal rats which were subjected to unilateral common carotid artery occlusion and exposed to hypoxic condition for 3 hours. The protective effect of XO inhibitor was examined by the cell number, cell viability, LDH and PS on 14 days after H-I with allopurinol intraperitoneal injection 15 minutes prior to H-I. In addition, the effect of allopurinol on PKC activity in hypoxic conditions was examined in neurons. Results : 72 hours from H-I, the cell numbers and viability were decreased significantly in time-dependent manner on neurons and those of astrocytes also decreased slightly, compared with control. In neonatal rats treated with H-I, the cell number, cell viability, and PS in neurons were decreased, but LDH was increased significantly compared with control. In neonatal rats pretreated with allopurinol, the cell number and viability, and PS in neurons were increased and LDH was decreased significantly compared with H-I. PKC was increased remarkably after hypoxic condition. But PKC was decreased significantly against hypoxic condition after allopurinol pretreatment. Conclusion : From these results, it is suggested that H-I is more toxic in neurons than astrocytes and allopurinol is very protective with increasing of PS, and decreasing of LDH and PKC in neurons from hypoxic-ischemic condition.
In order to develop liquid fertilizer using rendering by-product, rates and velocities of degradation from rendering by-product using pig cadaver investigated under different amount of injection and time with degradation agents (KOH, NaOH, $HNO_3$ and $H_2SO_4$). The amount of residue by degradation agent of $HNO_3$ treatment method was higher than that in KOH, NaOH and $H_2SO_4$ treatment methods. The degradation velocities (K; $hr^{-1}$) of rendering by-product in KOH treatment method were higher in the order of 25% ($0.0309hr^{-1}$) > 30% ($0.0268hr^{-1}$) > 20% ($0.0142hr^{-1}$) > 15% ($0.0111hr^{-1}$) > 10% ($0.0102hr^{-1}$) of weight of rendering by-product. In all conditions, the degradation velocity of rendering by-product using $H_2SO_4$ 30% of weight of rendering by-product was rapid than that for KOH, NaOH and $HNO_3$ treatment methods. Degradation rates of rendering by-product using NaOH were similar to that of KOH and $H_2SO_4$ except for $HNO_3$ under injecting 25% and 30% of rendering by-product weight.
This study was accomplished using attached $A^2/O$ process that contains nonsurface-modified and surface-modified polyethylene media inside the Anaerobic/Anoxic, Oxic tank, respectively. We could make the hydrophobic polyethylene media have hydrophilic characteristics by radiating ion beam on the surface of the media. The objectives of this study is to investigate the removal efficiencies of the organics and nitrogen when the step feed ratio of raw wastewater into anaerobic and anoxic tank is changed. In this case, we assumed that the denitrification rate can be improved because the nitrifiers in anoxic tank can perform denitrification using RBDCOD instead of artificial carbon sources (for example, methanol, etc.). The wastewater injection rate into anaerobic/anoxic tank was set up by the ratio of 10 : 0, 9 : 1, 8 : 2, 6 : 4, and the results of BOD removal efficiency showed similar trends with $93.3\%,\;92.6\%,\;92.4\%\;and\;91.6\%$, respectively. But the BOD removal efficiency (utilization of the organics) in the anoxic tank was in the order of 9 : 1 $(84.8\%)$, 10 : 0 $(77.0\%)$, 8 : 2 $(75.3\%)$, and 6 : 4 $(61.1\%)$. The T-N removal efficiency was most high when the ratio is 9 : 1 $(67.4\%)$, and other conditions, 10 : 0, 8 : 2, 6 : 4, showed $61.3(\%),\;60.7\%,\;55.5\%$, respectively; the ratio 6 : 4 was found to be lowest T-N removal efficiency, lower than the ratio 9 : 1 by $12\%$. Though the nitrification rate of the ratio 10 : 0, 9 : 1, and 8 : 2 showed similar levels, the ratio 6 : 4 showed considerable inhibition of nitrification, ammonia was the great portion of the effluent T-N. The advantages of this process is that this process is cost-saving, and non-toxic methods than injecting the artificial carbon source.
Kang, Chun Goo;Park, Hoon-Hee;Oh, Shin Hyun;Lee, Han Wool;Kim, Jung Yul;Oh, Joo Yung;Lee, Ju Young;Kim, Jae Sam;Lee, Chang Ho
The Korean Journal of Nuclear Medicine Technology
/
v.17
no.2
/
pp.3-9
/
2013
Purpose: Currently commercially available phantom can reproduce and evaluate only a static situation, the study is incomplete research on phantom and system which is can confirmed functional situation in the kidney by time through dynamic phantom and blood flow velocity, various difference according to the amount of radioactive. Therefore, through this study, it has produced the dynamic kidney phantom to reproduce images through the dynamic flow of the kidney, it desire to evaluate the usefulness of nuclear medicine imaging. Materials and Methods: The production of the kidney phantom was fabricated based on the normal adult kidney, in order to reproduce the dynamic situation based on the fabricated kidney phantom, in this study it was applied the volume pump that can adjust the speed of blood flow, so it can be integrated continuously radioactive isotopes in the kidney by using $^{99m}Tc-pertechnate$. Used the radioactive isotope was supplied through the two pump. It was confirmed the changes according to the infusion rate, radioactive isotopes and the different injection speeds on the left and right, analysis of the acquired images was done by drawn ten times ROI in order to check the reproducibility of each on the front and rear of the kidney and bladder. Results: Under the same conditions infusion rate 40 mL/min fixed to adjust the pressure of the pump when the radiopharmaceuticals between 2-3 minutes in the most integrated in the kidney phantom was excreted inthe bladder. Glomerular filtration rate (GFR), respectively, by each device SYMBIA 1,091 mL/min, FORTE 1,232 mL/min, ARGUS 1,264 mL/min, INFINIA 1,302 mL/min in that there isno statistically significant difference was found, Tmax values and T1/2 values stars from all equipment with no statistically significant difference was found. CV values of the coefficient of variation less than 5% was found to be repeatable, and to 2.67% of the lowest SYMBIA appeared, INFINIA was the highest in the 4.86%. Conclusion: Through this study, the results showed that the dynamic kidney phantom system is able to similarly reproduce renogram in the actual clinical. Especially, the depicted over time for the flow to be excreted through the kidney into the bladder was adequately reproduce, it is expected to be utilized as basic data to check the quality of the dynamic images. In addition, it is considered to help in the field of functional imaging and quality control.
This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.
Purpose: Recently, massive proteinuria has been observed in some transplant patients after switching cyclosporine A (CsA) to sirolimus. To evaluate the pathogenesis of sirolimus-associated proteinuria, we investigated the early changes in slit diaphragm molecules by various administrative conditions of sirolimus and CsA. Methods: In vitro-Mouse podocytes were incubated with buffer (C), sirolimus ($10\;{\mu}g/mL$) after CsA ($10\;{\mu}g/mL$) (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S) for 12, 24, and 48 hours. In vivo- twenty four SPF female Wistar rats were divided into 4 groups buffer (C), sirolimus after 2 weeks of CsA (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S). All groups were treated by intraperitoneal injection every other day for 4 weeks (CsA: 25 mg/kg, sirolimus: 0.5 mg/kg). The changes in mRNA of slit diaphragm molecules were examined by RT-PCR. Results: The mRNA of nephrin was significantly decreased in group C-S and C+S in vitro. In vivo, the mRNA of nephrin in all groups using sirolimus and the mRNA of podocin in group C-S and C+S were decreased. Microscopically, group C-S and C+S showed small vacuolization and calcification in proximal tubular epithelial cells. Immunohistochemistry using nephrin and podocin antibodies did not show remarkable decrease of staining along the glomerular capillaries. Electron-microscopically, focal fusion of foot processes was seen in group C-S and C+S. Conclusion: This study suggests the decrease of slit diaphragm molecules (nephrin and podocin) in podocyte may be one of the causes of sirolimus associated proteinuria, and podocyte injury by sirolimus may need a primary hit by CsA to develop the proteinuria.
Park, Seok-Gun;Lee, Yeon-Hee;Rhyu, Jin-Woo;You, Sun-Mee
The Korean Journal of Nuclear Medicine
/
v.32
no.2
/
pp.151-160
/
1998
Purpose: Piccolo et al. advocated the usefulness of $^{99m}Tc$-MDP breast scan in differential diagnosis of breast mass with high accuracy. But there are little or no follow-up studies about it as we know. We studied $^{99m}Tc$-MDP uptake pattern and lesion/background ratio in patients complaining palpable breast lesions to evaluate the clinical usefulness of $^{99m}Tc$-MDP scan. Materials and Methods: Total 34 patients were studied with physical examination, mammo-gram and $^{99m}Tc$-MDP scan prospectively. Anteroposterior and both lateral view of breast were obtained 5 minutes after iv injection of 740 MBq $^{99m}Tc$-MDP. Breast uptake pattern of $^{99m}Tc$-MDP was analyzed by a grade system: 0=no uptake, grade 1=bilateral diffuse uptake, grade 2=asymmetric faint uptake, grade 3=focal hot uptake. 20 cases were pathologically confirmed by excision biopsy or aspiration biopsy. 14 cases were normal in physical examination and mammogram. Results: Pathologic results showed 7 carcinomas, 6 benign solid tumors, and 7 fibrocystic changes. Grade 3 pattern of $^{99m}Tc$-MDP uptake was noted in 4/7 carcinomas, 3/6 benign solid tumors, and 1/7 fibrocystic changes. Grade 2 pattern was 217, 0/7, 3/7 respectively. The average L/B ratio was 1.66 in carcinomas, 1.68 in benign solid masses, 1.20 in fibrocystic diseases, 1.05 in normal patients. L/B ratio was higher in carcinoma and benign mass groups than in fibrocystic change and normal control groups(p=0.005). But there was no statistical difference between L/B ratio of malignant mass group and benign mass group. Conclusion: $^{99m}Tc$-MDP scan is not suitable to routine clinical use for breast mass diagnosis. It might be used in limited conditions when whole body bone scan is planned.
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