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Optimization of Retrovirus Mediated-Gene Transfer into Hematopoietic Stem Cells (Retrovirus를 이용하여 조혈모세포에 유전자를 전달하기 위한 최적화)

  • Kim, Sang-Gyeong;Seo, Heon-Seok;Lee, Jong-Won;Sin, Dong-Geon;Lee, Jae-Gwan;Kim, Hyeon-Min;Kim, Jae-Sik;Seo, Jang-Su
    • KSBB Journal
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    • v.14 no.5
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    • pp.593-599
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    • 1999
  • In this study, optimal conditions to infect CD34 positive cells containing hematopoietic stem cells obtained from cord blood and bone marrow were found using two different retroviral vectors expressing human growth hormone (hGH) and $\beta$-galactosidase. CD34 positive cells were successfully infected with recombinant retroviruses only when the CD34 positive cells were co-cultured with packaging cells secreting recombinant retroviruses. To find the highest infection efficiency for the gene transfer, CD34 positive cells from cord blood were co-cultured with packaging cells secreting recombinant retroviruses encoding E. coli lacZ gene. The highest infection efficiency was obtained when CD34 positive cells were cultured for 3 days, and then co-culturing was done for another 2 days. When CD34 positive cells from bone marrow were co-cultured with packaging cells secreting recombinant retroviruses encoding hGH gene, the maximum amount of hGH was also secreted at the same conditions found above, i.e. 3 days of culture and 2 days of co-culture. These results show that there are optimal conditions for the gene transfer into hematopoietic stem cells regardless of sources of target cells or retroviral vectors used to infect.

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Optimization of the Reaction Conditions and the Effect of Surfactants on the Kinetic Resolution of [R,S]-Naoroxen 2,2,2-Trifluoroethyl Thioester by Using Lipse (리파아제를 이용한 라세믹 나프록센 2,2,2-트리플로로에틸 씨오에스터의 Kinetic Resolution에서 반응조건 죄적화와 계면활성제 영향)

  • Song, Yoon-Seok;Lee, Jung-Ho;Cho, Sang-Won;Kang, Seong-Woo;Kim, Seung-Wook
    • KSBB Journal
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    • v.23 no.3
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    • pp.257-262
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    • 2008
  • In this study, the reaction conditions for lipase-catalyzed resolution of racemic naproxen 2,2,2-trilfluoroethyl thioester were optimized, and the effect of surfactants was investigated. Among the organic solvents tested, the isooctane showed the highest conversion (92.19%) in a hydrolytic reaction of (S)-naproxen 2,2,2-trifluoroethyl thioester. In addition, the isooctane induced the highest initial reaction rate of (S)-naproxen 2,2,2-trifluoroethyl thioester ($V_s=2.34{\times}10^{-2}mM/h$), the highest enantioselectivity (E = 36.12) and the highest specific activity ($V_s/(E_t)=7.80{\times}10^{-4}mmol/h{\cdot}g$) of lipase. Furthermore, reaction conditions such as temperature, concentration of the substrate and enzyme, and agitation speed were optimized using response surface methodology (RSM), and the statistical analysis indicated that the optimal conditions were $48.2^{\circ}C$, 3.51 mM, 30.11 mg/mL and 180 rpm, respectively. When the optimal reaction conditions were used, the conversion of (S)-naproxen 2,2,2-trifluoroethyl thioester was 96.5%, which is similar to the conversion (94.6%) that was predicted by the model. After optimization of reaction conditions, the initial reaction rate, lipase specific activity and conversion of (S)-naproxen 2,2,2-trifluoroethyl thioester increased by approximately 19.54%, 19.12% and 4.05%, respectively. The effect of surfactants such as Triton X-100 and NP-10 was also studied and NP-10 showed the highest conversion (89.43%), final reaction rate of (S)-naproxen 2,2,2-trifluoroethyl thioester ($V_s=1.175{\times}10^{-2}mM/h$) and enantioselectivity (E = 59.24) of lipase.

Effect of Genistein on the Onset of Puberty in Female Rats (암컷 흰쥐의 사춘기 개시에 미치는 Genistein의 효과)

  • Lee, Kyeung-Yeup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.10 no.1
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    • pp.55-61
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    • 2006
  • There is growing concern that dietary soy intake is associated with protection of breast cancer. However, questions persist on the potential adverse effects of the main soy constituent genistein(GS) on female reproductive physiology. In this study, we examined whether prepubertal exposure to GS affected on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. GS(100mg/kg/day) was administrated daily from postnatal day 25(PND 25) to the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the day after VO occurred. Gross anatomy and tissue weight were compared to test the GS's effect on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in tissues. Specific radioimmunoassay(RIA) were carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptors(PR), total RNAs were extracted from ovary and uterus and were applied to semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a results, advanced VO was shown in the GS group(PND $31.2{\pm}0.6$) compared to the vehicle group (PND $35.3{\pm}0.7$). GS treatment significantly increased wet weight of ovaries and uteri compared to the vehicle group. Increased serum LH levels were also shown in the GS group. Graafian follicles and corpora lutea(CL) were observed only in the ovaries from GS treated animals. Similarly, hypertrophy of luminal and glandular uterine epithelium were found only in the GS group. Collectively, these effects were probably due to the estrogenic effects of GS. In the semi-quantitative RT-PCR studies, the transcriptional activities of PR in both ovary and uterus from GS-treated group were significantly higher than those from the vehicle group. The present studies demonstrated that acute exposure to GS, at levels comparable to the ranges of human exposure, during the critical period of prepubertal stage activates the reproductive system resulting precocious puberty in immature female rats.

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Investigation of Helicobacter pylori cagE Locus Diversity on Interleukin-8 Induction in AGS Cell-line (Helicobacter pylori cagE 유전자 다형성이 위상피세포주에서 Interleukin-8 유도능에 미치는 영향)

  • Lee Su-Min;Lee Hak-Sung;Lee In-Suk;Co Yu-Kyung;Han Hae-Won;Oh Jung-Hwan;Park Jae-Myung;Choi Myung-Gyu;Chung In-Sik
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.906-912
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    • 2004
  • Helicobacter pylori infection is highly prevalent, as high as 2/3 of whole population infected, in Korea. H. pylori infection initiates inflammation by induction of interleukin-8 through type IV secretion of CagA. It was recently suggested that induction failure of IL-8 is not associated with defect in cag PAI but associated with cagE locus diversity. This study was designed to investigate ability of 11-8 in-duction according to sequence variation within the cagE gene, cagA TP motifs and vacA m-types in vitro study using AGS cell-line, and to evaluate its association with different clinical outcome. Seventy-four H. pylori stains were isolated from 23 patients with gastric cancer (Ca), 24 subjects with gastritis (G) and 27 patients with duodenal ulcer (Du) in Kangnam St. Mary's Hospital, Seoul, Korea. cagE gene diversity was confirmed by the PCR-RFLP methods with MboI/NlaIII and tyrosine phosphate motifs (TPMs) of cagA was determined TPM-A and C by using DdeI/Tsp5091 restriction enzyme and TPM-B was determend by Real time PCR the method of Owen et al. and IL-8 was measured by ELISA assay. IL-8 activity was positively detected in 59 among 74 strains $(79.7\%)$. IL-8 secretion was significantly increased in MboI A and MboI B type compared to MboI C type and in MboI/NlaIII A-C and B-C type than C-C type. 1L-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. There was no significant difference in IL-8 activity among patient groups. cagE gene diversity is thought to be mainly associated with the induction of IL-8 in H. pylori infection.

Effect of Atmospheric Pressure Plasma on the Quality of Commercially Available Sunsik (대기압 플라즈마가 선식의 품질 특성에 미치는 영향)

  • Kim, Hyun-Joo;Woo, Koan Sik;Jo, Cheorun;Lee, Seuk Ki;Park, Hye Young;Sim, Eun-Yeong;Won, Yong-Jae;Lee, Sang-Bok;Oh, Sea-Kwan
    • Journal of Food Hygiene and Safety
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    • v.31 no.5
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    • pp.375-379
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    • 2016
  • Atmospheric pressure plasma (APP) was applied to examine microbial safety and qualities of commercial Sunsik. APP was generated in a square-shaped plastic container (250 W, 15 kHz, ambient air) and dielectric barrier discharge plasma treatment was applied for periods of 0, 5, 10, and 20 minutes. The total aerobic bacterial count in the control was 4.44 log CFU/g. Under plasma treatment for 20 minutes, Sunsik samples inoculated with Bacillus cereus, B. subtilis, and Escherichia coli O157:H7 resulted in a reduction of bacterial counts by approximately 2.20, 2.22, and 2.52 log CFU/g, respectively. The pH of the sample was found to decrease after APP treatment. Although hunter color $L^*$ of Sunsik increased, $a^*$ and $b^*$ value decreased as a result of APP. Increasing the APP time also enhanced the peroxide value. Further, sensory evaluation revealed that APP decreased color, flavor, taste and overall acceptability. The results of this study indicated that APP treatment improved the microbial quality of Sunsik, although further studies should be conducted to reduce the deterioration of sensory quality induced by APP.

A Study on Evaluating Solute Excretion in the Normal Neonate (정상 신생아에서 용질배설 측정 의의에 관한 연구)

  • Choi Jeong Hoon;Kim Mi Kyung;Yoo Kee Hwan;Hong Young Sook;Lee Joo Won;Kim Soon Kyum
    • Childhood Kidney Diseases
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    • v.4 no.1
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    • pp.6-10
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    • 2000
  • Purpose: To evaluate whether the urinary creatinine concentration is a reliable reference value to standardize urinary solute excretion in a spot urine sample during the first week of life. Methods: Spontaneously voided urine specimens were obtained in 49 healthy full term neonates, and in 33 healthy older children with the median ages of $5.7{\pm}4.3$ years, two urine samples were available with an interval of 2 to 3 days. Urine creatinine concentration was determined by the Jaffe test(CoBAS, Integra, Roche, Swiss). Uurine osmolality was determined by the freezing point depression test(Multi-osmette, Precision, USA). Results: Mean urinary creatinine and osmolality values of the first urine samples were not significantly different with the second urine samples in each group. Mean urinary creatinine and osmolality values in neonates were significantly different from the older children of the each urine sample(P<0.01). In neonates, the mean of the urinary oreatinine/osmolality ratios was higher than that of the older children(P<0.01). The urinary creatinine and the creatinine/osmolality values of the first urine samples were closely correlated with those of the second samples in both two groups(P<0.001). Conclusion: The urinary creatinine concentration during the first day of life is relatively stable, even when corrected for urinary osmolality The urinary creatinine and the urinary creatinine/osmolality ratio, therefore, can be used to standardize the urinary excretion of solutes in the neonate.

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Effect of the Various Sources of Dietary Additives on Growth, Body Composition and Shell Color of Abalone Haliotis discus hannai (다양한 원료의 사료첨가제가 전복의 성장, 체조성 및 패각 색채에 미치는 영향)

  • Cho, Sung-Hwoan;Park, Jung-Eun;Kim, Chung-Il;Yoo, Jin-Hyung;Lee, Sang-Min;Choi, Cheol-Young
    • Journal of Aquaculture
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    • v.19 no.4
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    • pp.275-280
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    • 2006
  • Effect of the various sources of dietary additives on growth, body composition and shell color of abalone Haliotis discus hannai was investigated for 16 weeks. Forty juvenile abalone averaging 13.5 g were randomly stocked into 21 of 50 L plastic rectangular containers each. Eight kinds of additives were prepared for this study: four commercially available microalgae [Haeatococcus (Hae), Isochrysis galbana (Iso), Shizochytrium (Sch) and Spirulina (Spi)], three crustacean meals [krill meal (KM), shrimp head meal (Shm) and red crab meal (Rcm)], and green tea by-product (Gre). In addition, dry sea tangle (Dst), Laminaria japonica, as a control, was prepared. Casein, dextrin and a mixture corn oil and fish oil was protein, carbohydrate and lipid sources, respectively, in the experimental diets. The 2% each additive was included into the experimental diets. The experimental diets were fed to abalone once a day at the ratio of $1.5{\sim}2.0%$ total biomass of abalone with a little leftover throughout the 16-week feeding trial. Survival of abalone was not significantly (P>0.05) affected by the experimental diets. However, weight gain of abalone fed the all experimental diets containing the various sources of additives was significantly (P<0.05) higher than that of abalone fed the Dst diet. Weight gain of abalone fed the Spi diet was highest and Shi, KM and Iso diets in order. Shell length and the ratio of soft body weight to body weight of abalone was not significantly (P>0.05) affected by the experimental diets. However, shell width of abalone fed the all experimental diets containing the various sources of additives was significantly (P<0.05) higher than that of abalone fed the Dst diet. The shell color of abalone fed the Spi diet was improved the most distinctively and similar to that of natural abalone. Therefore, it can be concluded that the experimental diets with the various sources of additives (microalgae and crustacean meals) was effective to improve growth of abalone and dietary inclusion of Spirulina was most effective to improve shell color of abalone.

Clinical and Bacteriologic Efficacy of Cefdinir on Pharyngitis and Pharyngotonsillitis Caused by Group A Beta Hemolytic Streptococci in Children (Group A-beta Hemolytic Streptococci에 의한 소아 인두편도염에 있어서 Cefdinir의 항균력과 임상적 및 세균학적 효과에 관한 연구)

  • Chung, Ji-Young;Sin, Seon-Hee;Ahn, Young-Min;Ahn, Byung-Moon;Sin, Young-Gyu;Bae, Young-Min;Park, Soo-Eun;Kim, Jong-Guk;Lee, Jong-Guk;Ma, Saung-Hyuk;Chang, Jin-Kun;Cha, Sung-Ho
    • Pediatric Infection and Vaccine
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    • v.10 no.1
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    • pp.95-101
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    • 2003
  • Objective : To determine the clinical and bacteriologic efficacy and safety of Cefdinir in acute pharyngitis and pharyngotonsillitis caused by group A beta hemolytic streptococci in pediatric patients. Methods : Children aged 3 through 12 years who visited the hospitals enrolled in this study with signs and symptoms of pharyngitis or pharyngotonsillitis since May to December 2002, were taken throat culture and given Cefdinir(12 mg/kg/day, in three divided doses) for 7 days. Two hundred thirty five patients were enrolled and 90 patients who showed positive culture results were followed up for the signs and symptoms during the treatment to determine clinical efficacy. Follow-up culture were done at the end of the study and bacteriologic efficacy was determined Results : Ninety out of 235 patients who visited the hospitals with the signs and symptoms of pharyngitis showed positive growth on throat culture. Seventy nine patients were clinically and microbiologically assessable. The bacteriologic eradication rates of S. pyogenes were 100% in all the children treated with Cefdinir. Clinical cure rates were not different between less than 7 days-treated group(75%) and just 7 days-treated group(98.6%)(P=0.077). Two patients reported adverse reaction during Cefdinir treatment. Antimicrobial sensitivity of Cefdinir against S. pyogenes was 100% with range of MIC being less than 0.5 ${\mu}g/mL$. Conclusion : It seems that Cefdinir is one of reliable and well-tolerated drugs for the treatment of group A beta hemolytic streptococcal pharyngotonsillitis in children. It needs to be investigated short term efficacy in terms of improving drug compliance and impact of economic point of view. We can recognized that still high rate of the erythromycin resistant group A streptococci in our community should be monitored every year.

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A Study on the Protective Effects of Glutathione on Cytotoxicity of Mercury and Cadmium (수은 및 카드뮴의 세포독성에 대한 Glutathione의 역할에 관한 연구)

  • Jeong, Jae-Ho;Kim, Jun-Youn;Koh, Dai-Ha
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.2
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    • pp.170-176
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    • 1999
  • Objectives: To evaluate the protective effects of glutathione (GSH) on the cytotoxicity of mercurial compounds$(CM_3HgCl,\;HgCl_2)$ or cadmium chloride$(CdCl_2)$ in EMT-6 cells. Methods: The compounds investigated were $CH_3HgCl,\;HgCl_2,\;CdCl_2$, GSH, buthionine Sulfoximine(BSO), L-2-oxothiazolidine-4-carboxylic acid(OTC). Cytotoxicity analysis consist of nitric oxide(NO) production, ATP production and cell viability. Results: Mercurial compounds and cadmium chloride significantly decreased cell viability and the synthesis of NO and cellular ATP in EMT-6 cells. GSH was not toxic at concentrations of 0-1.6 mM. In the presence of GSH, mercurial compounds and cadmium did not decrease the production of ATP and nitrite in EMT-6 cells. The protective effects of GSH against the cytotoxicity of mercurial compounds and cadmium depended on the concentration of added GSH to the culture medium for EMT-6 cells. We evaluated the effects of intracellular GSH level on mercury- or cadmium-induced cytotoxicity by the pretreatment experiments. Pretreatment of GSH was not changed ${NO_2}^-$ and ATP production, and pretreatment of BSO was decreased in dose and time-dependent manner. Pretreatment of OTC was increased ${NO_2}^-$ and ATP production in dose- and tine-dependent manner. Because intracellular GSH level was increased by OTC pretreatment, the protective effect on mercury- and cadmium-induced cytotoxicity was increased. Conclusions: These results indicated that sulfhydryl compounds had the protective effects against mercury-induced cytotoxicity by the intracellular GSH levels.

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Biological Control of Fusarium Stalk Rot of Maize Using Bacillus spp. (Bacillus spp.를 이용한 옥수수 밑둥썩음병의 생물학적 방제)

  • Han, Joon-Hee;Park, Gi-Chang;Kim, Joon-Oh;Kim, Kyoung Su
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.280-289
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    • 2015
  • Maize (Zea mays L.) is an economically important crop in worldwide. While the consumption of the maize is steadily increasing, the yield is decreasing due to continuous mono-cultivation and infection of soil-borne fungal pathogens such as Fusarium species. Recently, stalk rot disease in maize, caused by F. subglutinans and F. temperatum has been reported in Korea. In this study, we isolated bacterial isolates in rhizosphere soil of maize and subsequently tested for antagonistic activities against F. subglutinans and F. temperatum. A total of 1,357 bacterial strains were isolated from rhizosphere. Among them three bacterial isolates (GC02, GC07, GC08) were selected, based on antagonistic effects against Fusarium species. The isolates GC02 and GC07 were most efficient in inhibiting the mycelium growth of the pathogens. The three isolates GC02, GC07 and GC08 were identified as Bacillus methylotrophicus, B. amyloliquefaciens and B. thuringiensis using 16S rRNA sequence analysis, respectively. GC02 and GC07 bacterial suspensions were able to suppress over 80% conidial germination of the pathogens. GC02, GC07 and GC08 were capable of producing large quantities of protease enzymes, whereas the isolates GC07 and GC08 produced cellulase enzymes. The isolates GC02 and GC07 were more efficient in phosphate solubilization and siderophore production than GC08. Analysis of disease suppression revealed that GC07 was most effective in suppressing the disease development of stalk rot. It was also found that B. methylotrophicus GC02 and B. amyloliquefaciens GC07 have an ability to inhibit the growth of other plant pathogenic fungi. This study indicated B. methylotrophicus GC02 and B. amyloliquefaciens GC07 has potential for being used for the development of a biological control agent.