• Title/Summary/Keyword: Cell treatment

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Characterization and Cloning of the Gene Encoding Autoregulator Receptor Protein from Streptomyces longwoodensis (Streptomyces longwoodensis로부터 Autoregulator Receptor Protein 유전자의 클로닝 및 특성)

  • Yeo Soo-Hwan;Lee Sung-Bong;Kim Hyun-Soo
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.96-105
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    • 2005
  • For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

Study on the Genetic Characteristics of Waterlogging Tolerant Pepper (Capsicum annuum L.) for Breeding Tolerant Varieties against Flooding Stress (내습성 고추 품종 육성을 위한 선발계통의 유전적 특성 구명)

  • Yang, Eun Young;Chae, Soo-Young;Hong, Jong-Pil;Lee, Hye-Eun;Park, Eun Joon;Moon, Ji-hye;Park, Tae-Sung;Roh, Mi-Young;Kim, Ok Rye;Kim, Sang Gyu;Kim, Dae Young;Lee, Sun Yi;Cho, Myeong Cheoul
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1111-1120
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    • 2017
  • This study was conducted to select pepper lines that were tolerant to excessive water injury among the pepper germplasm and investigate the genetic characteristics of those lines to contribute to the breeding of pepper cultivars with stable productivity in abnormal weather. Each of the tolerant and susceptible lines went through immersion treatment, and differentially expressed genes between them were analyzed. The tolerant line showed increased expression of the CA02g26670 gene, which is involved in the CONSTANS protein pathway and regulation of flowering by day length, but it exhibited decreased expressions of CA01g21450, CA01g22480, CA01g34470, CA02g00370 and CA02g00380. The susceptible line showed increased gene expressions of CA02g09720, CA02g21290, CA03g16520, CA07g 02110, and CA12g17910, which are involved in the inhibition of proteolytic enzyme activity, DNA binding, inhibition of cell wall-degrading enzyme, and inhibition of nodulin, respectively. Meanwhile the expressions of CA02g02820, CA03g21390, CA06g17700 and CA07g18230 decreased in the susceptible line, in relation to calcium-ion binding, high temperature, synthesis of phosphocholine and cold stress, respectively. The expressions of genes related to apoptosis and peroxidase increased, while that of CA02g16990, which functions as a nucleoside transporter, decreased in both the tolerant and susceptible lines. Based on the different gene expressions between the tolerant and susceptible lines, further studies are needed on breeding abiotic stress-tolerant lines.

Optimization of Culture Conditions and Encapsulation of Lactobacillus fermentum YL-3 for Probiotics (가금류 생균제 개발을 위한 Lactobacillus fermentum YL-3의 배양조건 최적화 및 캡슐화)

  • Kim, Kyong;Jang, Keum-Il;Kim, Chung-Ho;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.255-262
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    • 2002
  • This experiment was performed to improve the stability of Lactobacillus fermentum YL-3 as a poultry probiotics. The culture conditions that improve acid tolerance of L. fermentum YL-3 were investigated by changing several factors such as medium composition, temperature, anaerobic incubation and culture time. Also, L. fermentum YL-3 was encapsulated with alginate, calcium chloride and chitosan. The stable culture conditions of L. fermentum YL-3 were obtained in anaerobic incubation using MRS media without tween 80 for 20 hour at $42^{\circ}C$. The capsule after treatment with 1% chitosan was formed close membrane by a bridge bond. Immobilization of L. fermentum YL-3 in capsule was observed by confocal laser scanning microscopy, and cell viability was $2.0{\times}10^9\;CFU/g$ above the average. L. fermentum YL-3 capsule after acid treated at pH 2.0 for 3 hour survived about 40%, but those encapsulated with 1% chitosan survived about 65%. Survival rate of capsule stored at room temperature decreased about $2{\sim}3$ log cycle during 3 weeks, but viability of capsule stored at $4^{\circ}C$ during 3 weeks maintained almost $10^8\;CFU/g$ levels.

Improvement of L-Lysine Productivity by Using Cell Fusion and Immobilized System (세포융합과 고정화 시스템을 이용한 L-Lysine의 생산성 향상)

  • Ryu, Beung-Ho;Kim, Hye-Sung;Roh, Myung-Hoon;Park, Bob-Gyu;Chung, Jong-Soon;Bai, Ki-Chul
    • Korean Journal of Food Science and Technology
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    • v.21 no.1
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    • pp.154-163
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    • 1989
  • This studies were designed to improve the productivity of L-lysine by protoplast fusion and immobilized system of fusants using strains of Brevibacterium flavum ATCC 21528, Brevibacterium lactofermentum ATCC 21086 and Corynebacterium glutamicum 820. Mutants were isolated with concentration method of $300{\mu}g/ml$ penicillin-G after treatment of $250{\mu}g/ml$ N-methyl-N-nitro-N-nitrosoguanidine. B. flavum $37-2(Hos^-,\;Kan^r,\;AEC^r)$, B. lactofermentum $6-2(Ile^-,\;Val^-,\;Str^r,\;AEC^r)$ and C. glutamicum 57-5$(Met^-,\;Thr^-,\;Rif^r,\;AEC^r)$ were isolated from mutants. Protoplasts were induced by being incubated with $500{\mu}g/ml$ lysozyme of lysis solution for 6 hr and the ratio of protoplast formation and regeneration were ranging from 97-99% and 33-37%, respectively. Fusion frequencies of fusants of BBFL 21, BCFG 37 and BCLG 59 were shown in the range from $1.25{\times}10^{-6}\;to\;5.83{\times}10^{-7}$ under the optimum conditions. The fusant BBFL 21 showed the highest productivity of $411.1\;ng/ml{\cdot}hr$ L-lysine in the lysine productivity broth at $30^{\circ}C$ for 72hr. In the immobilization systems, fusant BBFL 21 was employed in various polymer matrices such as sodium alginate, polyacrylamide, agar and ${\alpha}-carrageena$. The immobilization of sodium alginate showed the highest productivity of $413\;ng/ml{\cdot}hr$ L-lysine in the batch system. Continuous fermentation of immobilization system by using tube fermentor was produced the highest productivity $416.7\;ng/ml{\cdot}hr $ L-lysine under optimum condition.

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Effect of Fermented Cirsium japonicum Extract on Testosterone Deficiency Syndrome (엉겅퀴 발효 추출물을 통한 남성 갱년기 증상 개선 효과)

  • Jeong, Byung Seo;Kim, Seong Hoon;Kim, Hyun Pyo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.7
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    • pp.790-800
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    • 2017
  • As men get older, total testosterone levels decline gradually, and concentrations of free and bioavailable testosterone decline sharply with each decade beyond their 30s. Andropause or testosterone deficiency syndrome (TDS) is defined as a decrease in sexual satisfaction or decline in general well-being accompanied by low levels of testosterone in older men. This male climacteric is characterized by nervousness, reduced potency, decreased libido, irritability, fatigue, depression, memory problems, sleep disturbances, and hot flushes. Cirsium japonicum (CJ) is used as a traditional medicine for hemorrhage, blood congestion, and inflammation in Korea. However, there is no report on the efficacy of CJ treatment for TDS. In this study, we observed the mitigating effect of CJ extract (CE) and fermented CJ extract (FCE) on symptoms of TDS. In elderly male rats, total and testosterone levels, hind limbs muscles, forced swimming time, and total and motile sperm counts significantly increased after daily intake of CE and FCE for 6 weeks. In contrast, sex hormone binding globulin, retroperitoneal fat, total serum cholesterol, and triglyceride levels were significantly reduced in CE and FCE groups. However, there was no difference in prostate specific antigen, aspartate aminotransferase, and alanine aminotransferase levels among all groups, which means CE and FCE did not have putative adverse effects. In a cell experiment, we also observed that CE and FCE enhanced expression of genes related to testosterone biosynthesis but reduced genes involved in testosterone conversion. On the whole, these positive effects on TDS were greater in FCE compared to CE. Thus, these results suggest the potential of FCE as a promising natural product for recovering testosterone levels and alleviating TDS.

The Effect of Surfactant Therapy for Acute Lung Injury Induced by Intratracheal Endotoxin Instillation in Rats (기관내 내독소 투여로 유발된 흰쥐의 급성폐손상에서 surfactant의 치료효과)

  • Kang, Yun-Jung;Park, Yong-Bum;Jee, Hyun-Suk;Choi, Jae-Chol;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung-Whui
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.4
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    • pp.487-499
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    • 2000
  • Background : Acute lung injury is an hypoxic respiratory failure resulting from damage to the alveolar-capillary membrane, which can be developed by a variety of systemic inflammatory diseases. In this study the therapeutic effects of intra-tracheal pulmonary surfactant instillation was evaluated in the intratracheal endotoxin induced acute lung injury model of a rat. Methods : Twenty Sprague-Dawley rats were divided into 4 groups, and normal saline (2 ml/kg, for group 1) or LPS (5 mg/kg, for group 2, 3, and 4) was instilled into the trachea respectively. Either normal saline (2 ml/kg, for group 1 & 2, 30 min later) or bovine surfactant (15 mg/kg, 30 min later for group 3, 5 hr later for group 5) was instilled into the trachea. The therapeutic effect of intratracheal surfactant therapy was evaluated with one chamber body plethysmography (respiratory frequency, tidal volume and enhanced pause), ABGA, BAL fluid analysis (cell count with differential, protein concentration) and pathologic examination of the lung. Results : Intratracheal endotoxin instillation increased the respiration rate decreased the tidal volume and int creased the Penh in all group of rats. Intratracheal instillation of surfactant decreased Penh, increased arterial oxygen tension, decreased protein concentration of BAL fluid and decreased lung inflammation at both times of administration (30 minute and 5 hour after endotoxin instillation). Conclusion : Intratracheal instillation of surfactant can be a beneficial therapeutic modality as discovered in the acute lung injury model of rats induced by intratracheal LPS intillation. It deserves to be evaluated for treatment of human acute lung injury.

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Comparison of Size Criteria in Mediastinal Lymph Node Involvement of Adenocarcinoma of Lungs (폐 선암의 종격동 림프절 전이에 있어서 림프절 크기 기준의 비교)

  • Gu, Ki-Seon;Kuk, Hiang;Koh, Hyeck-Jae;Yang, Sei-Hun;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.4
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    • pp.542-547
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    • 1999
  • Background: Decision in mediastinal lymph node involvement of lung cancer by CT scan is very important and valuable for the treatment planning and prognosis prediction. In general, long diameter of mediastinal lymph node more than 15mm is used as criterion of lung cancer involvement. Adenocarci-noma has a tendency of early distant metastasis and micrometastasis, so adenocarcinoma may involve lymph node earlier and cannot be detected before lymph nodes are enlarged enough. The authors tried to determine the difference between two size criteria(15mm, 10mm) in adenocarcinoma for the detection of cancer involvement. Methods: Numbers of sample are 60 cases(male 46, female 14, median age: 61.5 years). According to pathology, squamous cancer 41, large cell cancer 2, adenocarcinoma 17. According to TNM stage, I 23, III 24, IIIA 13. Results : Mean long diameter of lymph node involvement is 16.0($\pm8.0$) mm in non-adenocarcinoma group, and that of adenocarcinoma group is 12.0($\pm3.2$) mm(p<0.05). If long diameter of lymph node larger than 15mm as involvement criterion is applied, sensitivity, specificity, positive predictive index, negative predictive index, accuracy of nonadenocarcinoma group are 54%, 100%, 100%, 83%, 86%, and those of adenocarcinoma group are 43%, 90%, 75%, 69%, 71%. If long diameter of lymph node larger than 10mm as involvement criterion is applied, sensitivity, specificity, positive predictive index. negative predictive index. accuracy of nonadenocarcinoma group are 65%, 77%, 61%, 92%, 79%, and those of adenocarcinoma group are 100%, 80%, 78%, 100%, 88%. Conclusion: Long diameter of lymph node larger than 10mm is more valuable criterion as lymph node involvement in adenocarcinoma of lungs.

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Factors Associated with the Development of Pleural Thickening in Tuberculous Pleurisy (결핵성 흉막염 치료 후 흉막비후의 예측인자)

  • Park, Jae-Seuk;Chun, Yong;Choi, Eun-Kyung;Jee, Young-Koo;Lee, Kye-Young;Kim, Keum-Youl
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.1
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    • pp.17-24
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    • 1999
  • Background : A sizable percentage of tuberculous pleurisy patients are known to have residual pleural thickening(RPT) despite adequate anti-tuberculous chemotherapy. But, the predictive factors related to the development of RPT is not well known. Therefore, we studied to determine which factors are related to the development of RPT after completion of therapy. Methods: By retrospective review of medical records, fifty-eight patients initially diagnosed as having tuberculous pleurisy between March 1995 and January 1998 were separated into two groups : 27 patients in group 1 had RPT on simple chest radiography, while 31 patients in group 2 had no RPT after 6 month of anti-tuberculous chemotherapy. The clinical characteristics, radiologic findings and pleural fluid findings of the two group were compared at the time of diagnosis and during the course of therapy. Results: 47% of patients had RPT after 6 month of chemotherapy, and RPT was more common in man than in women(54% vs 29%, p=0.092). In group 2 patients, complete resorption of pleural lesion occurred rather late stage of therapy(1-2 month: 26%, 3-4 month: 29%, 5-6 month: 45%). Group 1 patients had increased percentage of loculated pleural lesion(26 % vs 19%) and increased white blood cell and lymphocyte count, lactate dehydrogenase level in pleural fluid ($3527\pm5652$ vs $2467\pm2201$/ml, $2066\pm2022$ vs $1698\pm1835$/ml and $1636\pm1143$ vs $1441\pm923$IU/mL respectively) than group 2 at the time of diagnosis, but statistically insignificant. Duration of symptom prior to treatment, size of pleural effusion, presence of parenchymal lung lesion, level of total protein, glucose and adenosine deaminase(ADA)activity in pleural fluid were similar in both group. Conclusion: 53% of tuberculous pleurisy patients showed slow but complete resorption of pleural lesion after 6 month of chemotherapy. But, no clinical, radiological and pleural fluid findings are predictive for the development of RPT.

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Postoperative Radiation Therapy in the Management of Early Cervical Cancer (초기 자궁경부암의 수술 후 방사선치료의 효과)

  • Kim, Jae-Chul
    • Radiation Oncology Journal
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    • v.24 no.3
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    • pp.164-170
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    • 2006
  • [ $\underline{Purpose}$ ]: This study identified the result of postoperative radiation therapy and the prognostic factors to affect survival rates in cancer patients. $\underline{Materials\;and\;Methods}$: One hundred and thirty three patients with cervical cancer who were treated with postoperative radiation therapy following surgery at our institution between June 1985 and November 2002 were retrospectively analyzed. One hundred and thirteen patients had stage IB disease, and 20 patients had stage IIA disease. Histological examination revealed 118 squamous cell carcinoma patients and 15 adenocarcinoma patients. Sixty seven patients were noted to have stromal invasion greater than 10 mm, and 45 patients were noted to have stromal Invasion 10 mm or less. Positive lymphovascular invasion was found in 24 patients, and positive pelvic lymph nodes were noted in 39 patients. Positive vaginal resection margin was documented in 8 patients. All of the patients were treated with external beam radiation therapy to encompass whole pelvis and primary surgical tumor bed. Intracavitary radiation therapy was added to 19 patients who had positive or close surgical margins. $\underline{Results}$: Actuarial overall and disease-free survival rates for entire group of the patients were 88% and 84% at 5 years, respectively. Five-year disease-free survival rates for patients with stromal invasion greater than 10 mm and 10 mm or less were 76% and 97%, respectively (p<0.05). Also there was a significantly lower survival in patients with positive pelvic lymph nodes compared with patients with negative pelvic lymph nodes (p<0.05). However, lymphovascular invasion, positive vaginal resection margins were not statistically significant prognostic factors. Addition of neoadjuvant chemotherapy or type of surgery did not affect disease-free survival. $\underline{Conclusion}$: Postoperative radiation therapy appears to achieve satisfactory local control with limited morbidity in cervical cancer patients with high pathologic risk factors. Distant metastasis was a dominant failure pattern to affect survival in cervical cancer patients after radical surgery and radiation and more effective systemic treatment should be investigated in these high-risk patients.

Physicochemical properties of spray-dried rice flour with Lactobacillus plantarum CGKW3 (분무건조공정을 이용한 유산균포집 미분의 제조 및 물리화학적 특성)

  • Park, Hye-Mi;Lee, Dae-Hoon;Jeong, Yoo-Seok;Jung, Hee-Kyoung;Cho, Jae-Gon;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.22 no.3
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    • pp.392-398
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    • 2015
  • The physicochemical properties of spray-dried rice flour with Lactobacillus plantarum CGKW3 were investigated. Amylose and damaged starch contents of spray-dried rice flour (S10, S20, S30, and S50) with L. plantarum CGKW3 were 14.18~17.75% and 24.65~34.08%, respectively. The particle size of spray-dried rice flour was $82.28{\sim}131.17{\mu}m$. The rice flour with L. plantarum CGKW3 showed a good powder flowability. The water absorption and water solubility of spray-dried rice flour were 1.96~2.13 and 9.91~21.95%, respectively. Thermal properties measured by differential scanning calorimeter revealed that the enthalpy (${\Delta}H$) for starch gelatinization were highest in the rice flour (S50) with L. plantarum CGKW3. When compared, the viable cell number of spray-dried rice flour were found to be in the following order: S10 (5.78 log CFU/g) < S20 (6.38 log CFU/g) < S30 (6.69 log CFU/g) < S50 (7.11 log CFU/g). The survaival rate of L. plantarum CGKW3 was 60.02-73.85%, which reflected the improvement in the quality of rice flour with an increase in treatment concentration. Based on our results, spray-dried rice flour with L. plantarum CGKW3 could be used in various types of rice foods.