• Maxillofacial Plastic and Reconstructive Surgery
• /
• 제31권3호
• /
• pp.193-197
• /
• 2009

Background: Though it is clear that many types of viruses can infect the oral mucosa, its condition for infection is unclear. The purpose of this study was to analyze the conditions for viral infection of normal oral mucosa and explore the possibility of topical gene therapy to oral mucosa using a viral vector. Methods: Freshly taken fragments of the palate and the tongue of mice were used for organ culture. The specimens were exposed to green fluorescent protein (GFP)-adenoviral vector for 1 hour except for the control. Initial viral titer was $6.3{\times}10^{11}\;pfu/ml$ and the virus was diluted to working concentrations. The dilution ratio was 1:1,000 ($6.3{\times}10^8\;pfu/ml$), 1:10,000 ($6.3{\times}10^7\;pfu/ml$), and 1:100,000 ($6.3{\times}10^6\;pfu/ml$). They were then cultured on a stainless steel wire mesh in an organ culture dish. The specimens were stereoscopically examined every 24 hours for 6 days, after which they were fixed and analyzed through immunohistochemical methods Results: There was no visible expression in the control, $6.3{\times}10^6\;pfu/ml$, and $6.3{\times}10^7\;pfu/ml$ groups. Initial expression was observed at 24 hours after infection in both the palate and the tongue in $6.3{\times}10^8\;pfu/ml$ and the expression significantly increased until 3 days in the palate and 2 days in the tongue after infection (P<0.05). In both groups, the expression was mostly observed at the resection margin. Immunohistochemical studies showed that the epithelial cells were positive to GFP. Conclusion: The present study showed that topically applied adenovirus containing specific genetic information of GFP could successfully transduce GFP in normal oral epithelial cells at the resection margin in organ culture in terms of dose and exposure time.

• Asian-Australasian Journal of Animal Sciences
• /
• 제20권9호
• /
• pp.1407-1416
• /
• 2007

Ten, first lactation, 87.5%HF dairy cattle were used to investigate effects of long-term administration of recombinant bovine somatotropin (rbST) on nutrient uptake by the mammary gland at different stages of lactation. Measurements of arterial plasma concentrations and arterial-venous differences of metabolites across the mammary gland were performed in combination with measurment of mammary blood flow to estimate the mammary uptake. Animals in experimental groups were injected subcutaneously every 14 days from day 60 of lactation with a prolonged-release formulation of 500 mg of rbST (POSILAC, Monsanto, USA) or with sterile sesame oil without rbST in the control group. During early lactation, the milk yield of rbST-treated animals was higher than that of the control animals (p<0.05). The peak milk yield in both groups of animals declined from the early period of lactation with progression to mid- and late-lactation. No significant changes were observed in the concentration of milk lactose, while the concentrations of milk protein significantly increased as lactation advanced to mid- and late-lactation in both groups. Milk fat concentrations were significantly higher in rbST-treated animals than in control animals, particularly in early lactation (p<0.05). Mammary blood flow (MBF) markedly increased during rbST administration and was maintained at a high level throughout lactation. The mean arterial plasma concentrations for glucose and acetate of rbST-treated animals were unchanged. The net mammary glucose uptake of rbST-treated animals increased approximately 20% during early lactation, while it significantly decreased (p<0.05), including the arteriovenous differences (A-V differences) and extraction ratio across the mammary gland, as lactation advanced to mid- and late-lactation. A-V differences, mammary extraction and mammary uptake for acetate increased during rbST administration and were significantly higher (p<0.05) than in the control animals in early and mid-lactation. Mean arterial plasma concentrations for ${\beta}$-hydroxybutyrate and free glycerol were unchanged throughout the experimental periods in both groups. A-V differences and extraction ratio of ${\beta}$-hydroxybutyrate across the mammary gland did not alter during rbST administration. Mean arterial plasma concentrations for free fatty acids ($C_{16}$ to $C_{18}$), but not for triacylglycerol, increased in rbST-treated animals and were significantly higher than in control animals during early lactation (p<0.01). These findings suggest that an increase in MBF during rbST administration would not be a major determinant in the mediation of nutrient delivery and uptake by the mammary gland for increased milk production. Local changes in biosynthetic capacity within the mammary gland would be a factor in the utilization of substrates resulting in the rate of decline in milk yield with advancing lactation.

• 대한방사성의약품학회지
• /
• 제5권2호
• /
• pp.89-100
• /
• 2019

In our previous studies, we developed a ¹⁸F-labeled TSPO-binding ligand, named [¹⁸F]CB251, which has been proved to be a promising TSPO-binding PET radiotracer for the detection and monitoring of TSPO expression in pathological diseases. (Ki = 0.27 nM for TSPO, 1.96% ID/g of tumor uptake at 1h post-injection) Based on these results, we utilized 6,8-dichloro-2-phenylimidazo[1,2-a]pyridineacetamide analogs, CB185 (1) as a targeting moiety for the selective delivery of probes and anticancer molecules to TSPO-overexpressed tissues. In this study, we designed CB185 derivatives contains different PEG chains (n = 1, 3 and 5) and fluorescence dye (Cy5) to identify the necessary space between a TSPO-binding ligand and an anticancer agent. Three CB185 derivatives (11a-c) which contains Cy5 and PEG chain, were synthesized and the effect of PEG additive on their TSPO-binding affinities were evaluated using in vitro assays. The binding affinity for compounds 11a-c was lower than that of PK11195 (Ki = 3.2 nM), but still characterized by nanomolar binding affinity for TSPO (Ki = 46.5 nM for 11a, 51.0 nM for 11b, and 388.5 nM for 11c). These results showed that the conjugates are characterized by a moderate binding affinity toward TSPO except for compound 11c, which PEG chain consist of five PEG monomers. Our finding might add useful information to decide the appropriate PET chain length for developing new TSPO-targeting drug carriers.

• 한국가금학회지
• /
• 제35권3호
• /
• pp.241-245
• /
• 2008

형질 전환 닭 생산 방법들 가운데 목적 유전자 운반에 탁월한 능력이 있는 것으로 알려진 렌티바이러스는 배반엽 단계 수정란을 이용한 형질 전환 닭 생산 연구에 활발하게 이용되고 있다. 본 연구는 재조합 렌티바이러스를 이용하여 사람의 SOD-3 단백질이 닭의 ovalbumin 프로모터에 의해서 유도되는 형질 전환 닭을 생산하고자 하였다. 사람의 SOD-3 단백질은 호흡 과정에서 체내에서 생성되는 활성산소를 중화시키는 탁월한 기능이 있는 것으로 알려져 있다. 후보 병아리의 생산은 앞서 언급한 유전자를 가지는 $1{\times}10^6$ cfu/mL 재조합 렌티바이러스를 배반엽 단계 수정란의 미세 주입하고 대리난각 배양법을 이용하여 배양기에서 21일 동안 배양하는 방법으로 생산하였다. 유전자를 미세주입한 341개의 수정란에서 78수의 후보 형질 전환 병아리를 생산하였으며, 생산된 후보 형질 전환 병아리들의 유전 분석은 PCR 방법을 이용하여 검증하였다. 유전 분석 결과는 성 성숙에 이른 47수의 수컷들 가운데 2수의 정액에서 사람의 SOD-3 유전자가 존재함을 보였다. 이상의 연구 결과는 완전한 형태의 형질전환 닭 생산의 가능성을 보여주고 있다.

• Reproductive and Developmental Biology
• /
• 제30권2호
• /
• pp.87-92
• /
• 2006

The possibility of producing transgenic embryos expressing the green fluorescence protein (GFP) gene have been evaluated after transfer of exogenous gene into the porcine zygote cytoplasm using the intracytoplasm sperm injection (ICSI) as gene delivery method. For DNA binding to sperm heads, 0.05% Triton X-100 or Lipofectin was used. After injection of the sperm bound to DNA by means of Lipofectin or Triton X-100 triturate, the blastocyst formation rates on day 6 were not significantly different from that of ICSI only group (18.8, 19.2 and 25.3%). In terms of GFP expression, more embryos were in GFP form in Triton X-100 group than in Lipofectin group (40.6 vs 36.4%), while percentage of non-mosaic embryos expressing the GFP gene in all blastomere was higher (P<0.05) in Lipofectin group than in Triton X-100 group (4.2 vs 0.9%). ICSI embryos derived from sperm treated with Lipofectin/DNA complex was transferred into 3 recipients and were collected by uterine flushing on days 5, 7 and 15 after embryo transfer, and then GFP expression was observed by a fluorescence microscopy. Over 26% of the collected embryos were normally expressed GFP gene. These results suggest that foreign gene transfer method with DNA bound sperm caused minimal damage to structure of oocytes that can result to full development of porcine embryos. This was confirmed in this study when the embryos that were transferred after ISCI of DNA bound sperm had a normal development and gene expression until preimplantation.

• Journal of Nutrition and Health
• /
• 제36권7호
• /
• pp.736-742
• /
• 2003

Soybeans have been a major protein source for many centuries in Korea. Soybeans contain phytochemicals which are isoflavones, biochemically active component. Isoflavone is a kind of phytoestrogen, structurally and functionally similar to estrogen. It has been reported that the breast milk and blood of breast feeding mothers who consume soy products contain isoflavones. This study was conducted to investigate the effects of soy milk supplement on the isoflavones (daidzein, genistein) concentration of breast milk, plasma and urine from breast feeding woman. Seventeen healthy women who delivered at Kyung Hee Medical Center were recruited. For the first 2 weeks after delivery, seventeen women ingested 400 ml (isoflavone 43.2 mg) of soy milk on the given time starting from the day of giving birth. For the next 2 weeks, soy milk ingestion was withdrawn. Dietary intake and anthropometric data were checked and breast milk, blood, and 24 hr urine samples were collected on the day of giving birth, the 14th (the last day of the supplement phase) and 28th (the last day of the withdrawal phase) day, respectively. HPLC analysis was used to measure the concentration of isoflavones. Dietary intakes of the subjects were inadequate for the Korean RDA regardless of soy milk supplementation. Especially, intakes of vit A, calcium, and iron were very low. The Anthropometric data such as LBM, TBW, PIBW, BMI checked on the day of 14th decreased and maintained their levels by the 28th day. Daidzein concentration in breast milk was not affected by soy milk supplementation. However, genistein concentration decreased by the 28th day (14th day: 0.89 $\pm$ 0.10 $\mu$g/ml, 28th day : 0.48 $\pm$ 0.07 $\mu$g/ml) (p < 0.05). Plasma daidzein and genistein concentrations were not changed by the 14th day and decreased by the 28th day (14th day: 49.64 $\pm$ 3.30 ng/ml, 26.72 $\pm$ 2.90 ng/ml, 28th day: 38.30 $\pm$ 4.40 ng/ml, 6.51 $\pm$ 0.50 ng/ml, respectively) (p < 0.05). Twenty four hour urine concentrations of daidzein and genistein significantly increased by the 14th day and decreased by the 28th day (14th day: 5.80 :t 0.3 mg/d, 4.17 $\pm$ 0.2 mg/d, 28th day: 6.72 $\pm$ 0.4 mg/d, 5.09 $\pm$ 0.5 mg/d, respectively) (p < 0.001). The rate of urinary recovery of daidzein was greater than that of genistein. The results of this study indicate that the supplement of dietary soy milk to the lactating women elevates the contents of isoflavone in the breast milk.

• Journal of Yeungnam Medical Science
• /
• 제3권1호
• /
• pp.41-48
• /
• 1986

사람의 적혈구막으로부터 Band 3를 분리정제하고 이를 liposome 내로 도입시켜 그 결과를 관찰하였다. 적혈구를 약알칼리 저장액으로 용혈시켜 막을 분리한 후, 저이온강도 용액으로 처리하여 Band 4를 추출하였다. Triton X-100 추출액에 p-chloromercuribenzoate를 가하고 sucrose density gradient ultracentrifugation후 fractionation하여 Band 3를 정제하였다. phosphatidyl L-serine과 cholesterol을 1 : 1 molar ratio로 섞고 진공회전 증발기를 사용하여 chloroform을 제거한 후 Triton X-100을 제거한 Band 3용액을 가하고 sonication함으로 liposome(reverse-phase evaporation vesicle)을 만들면서 Band 3를 도입 시켰다 Band 3의 분리정제 및 liposome에 도입되었음은 sodium dodecyl sulfate-polyacrylamide gel 전기영 동 후 coomassie brilliant blue 염색으로 확인할 수 있었다.

• 대한영양사협회학술지
• /
• 제6권2호
• /
• pp.136-147
• /
• 2000

The aim of this research was to examine the management practices related to foodservice.clinical nutrition service provided by hospital foodservice operations. A survey of 30 hospital food nutrition service departments was undertaken and detailed information was collected from each, including surveys of 176 dietitians and 30 foodservice managers. Statistical data analysis was completed using the SAS/win 6.11 package for descriptive analysis and t-test. The results of this study can be summarized as follows : Out of 30 hospitals, 73.3%(22) were directly operated and 26.7%(8) were under contract foodservice management. Licensed number of beds were 768.7, with an average length of 11.4 days. The general characteristics of the dietitians were that 49.4% were aged between 25-29, and 60.8% were ordinary dietitians. 76.7% had bachelor's degree, 15.9% with master's degree and 7.4% were college graduates. Most hospital dietitians had internship training and 35.2% took a training course of 6 to 12 months. The average space of kitchen was 452.52 $m^2$, with 133.63$m^2$ for modified diet space and 18.13$m^2$ for nutrition counselling room space. The total number of normal meals was 1,255.47, with 502.93 of modified meals. The average calorie of normal meals was 2,145.04kcal, with 91.9g of protein contained in normal meal. The total food waste was 351.40kg. An average monthly cost for disposing food waste was 745,171.67 won. 83.3% of the kitchens were on the 1st basement and only 66.7%(20) of 30 hospitals had its own nutrition counselling room. 80% used dishwashers and 66.7% had an exclusive elevator for delivering meals. 80.0% of hospitals used centralized delivery service. 90% had established an area for hygiene division, 70% used dry zone within the dishwashers for sterilization, 66.7% provided spoon and chopstick, and 100% used dish cover. As means of food waste treatment, most hospitals(56.7%) used animal feed by contracters, followed by means of collection by contracters(30%) and in-house high speed fermentation machines(13.3%). It was found that only 33.3% hospitals regularly checked temperatures of the meals given to patients. Total productivity index was 3.72(meals/hour) in average, with an average productivity index for normal diet of 5.41. Average productivity index for modified diet was 4.62. Productivity indices for patient meals and clinical nutrition were 5.01(meals/hour) and 1.12(cases/hour) respectively and hospitals under self-operated foodservice management received higher points on clinical nutrition productivity index(P<0.01) than hospitals under contract foodservice management.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권17호
• /
• pp.7749-7754
• /
• 2015

Background: Currently, cationic liposome has become the commonly used vehicles for gene transfection. Furthermore, one of the most significant steps in microRNAs expression studies is transferring microRNAs into cell cultures successfully. In this study we aim to approach the feasibility of transfection of cervical cancer cell lines mediated by liposome and to obtain the optimized transfection condition for cervical cancer cell lines. Materials and Methods: $Lipofectamine^{TM}2000$ as the carrier, miR-101 mimic was transfected into Hela cells and Siha cells. Using green fluorescent protein as reporter gene, to set different groups according to cell seeding density, the amount of miRNA, miRNA and the proportion of Liposomes, Whether to add serum into medium to study their impact on the liposomal transfection efficiency. Finally, MTT assay was used to analyze the relative minimal cell toxicity of liposome reagents. Results: The seeding density of Hela cell line and Siha are $1.5{\times}10^4$ (per well of 24 well plates), miRNA amount is 1ul of both, the ratio of miRNA and liposome is 1:0.5 of Hela cell line; 1:0.7 of Siha cell line respectively, after 24 hours we can get the highest transfection efficiency. Compared with serum medium, only Siha cells cultured with serum-free medium obtained higher transfection efficiency before transfection (P<0.01). MTT assay showed that according to the above conditions which has the lowest cytotoxicity. Conclusions: The method of Liposome to transfected is a suitable way and it can be an efficient reagent for miRNA delivery for Hela cells and Siha cells in vitro. It may serve as a reference for the further research or application.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제35권2호
• /
• pp.112-119
• /
• 2009

Extracellular matrix(ECM) is known to function as a reservoir of endogenous growth factors, can be an effective delivery system of growth factor that easily lost bioactivity in solution. Fibrillar collagens like type I collagen, are the major constituent of the ECM and structural protein of bone. Also, it can be a scaffold for osteoblast migration. The purpose of this study was to compare the effects of absorbable Atelo-collagen Sponge($Teruplug^{(R)}$) insertion in tooth extraction sites on periodontal healing of the mandibular second molar after the extraction of the impacted third molar. The study population comprised 31 cases who had been scheduled for surgical removal of impacted mandibular third molars. All patients were in good general health and were not using any medication that would influence wound healing after surgery. In 15 cases control group, none was inserted into the tooth extraction site. In 16 cases experimental groups, $Teruplug^{(R)}$ was inserted into the tooth extraction site. We evaluated tooth mobility, pocket depth, gingival margin level preoperatively and 1 week, 2 weeks, 4 weeks, and 3 months postoperatively. The change was compared with two groups using Mann-Whitney test. The results were as follows. 1. There was no significant change of tooth mobility on both groups. 2. There was tendency of decreasing of previous pocket depth causing tooth extraction on both groups. 3. On gingival margin level, there was various change according to initial swelling and loss of attachment on both groups. 4. There was tendency of decreasing of gingival margin level on both groups because of removal of inflammation and decreasing of previous pocket depth. 5. There was large change of pocket depth on buccal middle, distal, lingual distal area because of tooth extraction and bone reduction. Compared with the control group and experimental group, we observed significant difference during some periods. The results of this study suggest that absorbable atelo-collagen sponge($Teruplug^{(R)}$) is relatively favorable bone void filler with prevention of tissue collapse, food packing and enhance periodontal healing.