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Development of succinate producing Cellulomonas flavigena mutants with deleted succinate dehydrogenase gene

  • Lee, Heon-Hak;Jeon, Min-Ki;Yoon, Min-Ho
    • Korean Journal of Agricultural Science
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    • v.44 no.1
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    • pp.30-39
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    • 2017
  • This study was performed to produce succinic acid from biomass by developing mutants of Cellulomonas flavigena in which the succinate dehydrogenase gene (sdh) is deleted. For development of succinate producing mutants, the upstream and downstream regions of sdh gene from C. flavigena and antibiotic resistance gene (neo, bla) were inserted into pKC1139, and the recombinant plasmids were transformed into Escherichia coli ET12567/pUZ8002 which is a donor strain for conjugation. C. flavigena was conjugated with the transformed E. coli ET12567/pUZ8002 to induce the deletion of sdh in chromosome of this bacteria by double-crossover recombination. Two mutants (C. flavigena H-1 and H-2), in which sdh gene was deleted in the chromosome, were constructed and confirmed by PCR. To estimate the production of succinic acid by the two mutants when the culture broth was fermented with biomass such as CMC, xylan, locust gum, and rapeseed straw; the culture broth was analyzed by HPLC analysis. The succinic acid in the culture broth was not detected as a fermentation products of all biomass. One of the reasons for this may be the conversion of succinic acid to fumaric acid by sdh genes (Cfla_1014 - Cfla_1017 or Cfla_1916 - Cfla_1918) which remained in the chromosomal DNA of C. flavigena H-1 and H-2. The other reason could be the conversion of succinyl-CoA to other metabolites by enzymes related to the bypass pathway of TCA cycle.

Psychosocial Assessment and Related Factors for Kidney Transplantation Candidates in South Korea: A Descriptive Correlational Study (한국 신장이식 대기자의 심리사회적 평가 및 영향 요인)

  • Chong, Hye Jin
    • Journal of Korean Biological Nursing Science
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    • v.21 no.4
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    • pp.249-258
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    • 2019
  • Purpose: The purpose of this study was to assess the psychosocial status of candidates for deceased donor kidney transplantation (DDKT) in South Korea and identify factors associated with high-risk psychosocial status. Methods: The descriptive cross-sectional design included application of a standardized transplantation-specific assessment tool, the Psychosocial Assessment Candidacy Transplantation (PACT) instrument, and review of candidates' medical records. A total of 157 patients awaiting DDKT were recruited from a Korean organ transplantation center. The collected data were analyzed using descriptive statistics and binary logistic regression. Candidates were divided into high- and low-psychosocial risk groups using 3 as the cutoff point of PACT. Results: The mean score for the PACT final rating was 2.10, and the lowest average subscale score was 2.30 for understanding the processes of transplantation and follow up. Poor physical candidacy, especially due to comorbidities, and older age were associated with high psychosocial risk. Conclusion: The results of this study showed that the psychosocial status of Korean DDKT candidates was problematic. Development of systematic management programs for effective clinical strategies in the care of candidates is crucial to maintain their desirable psychosocial status in case of an emergency DDKT operation and improve post-transplantation outcomes. Continuous education is absolutely necessary to enhance understanding of the transplantation process for high-risk psychosocial candidates. Additionally, nurses and transplantation professionals should screen candidates with high-risk psychosocial status early in the process using transplantation-specific assessment tools and provide intensive interventions, particularly for poor physical candidacy patients and older candidates.

Studies on Embryo Cryopreservation and Twinning by Embryo Transfer of Korean Native Cattle I. Transfer of Frozen-thawed Embryos and Production of Calves (한우 수정란의 동결보존 및 쌍자생산에 관한 연구 I. 동결 수정란의 이식과 자우 생산)

  • 손동수;김일화;이호준;서국현;이동원;류일선;이광선;전기준;손삼규
    • Journal of Embryo Transfer
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    • v.12 no.1
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    • pp.75-90
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    • 1997
  • This study was carried out to establish the techniques for producing the calves of genetically superior Korean Native cattle by transfer of frozen-thawed embryos. The effects of some factors related to embryo recovery following superovulation and pregnancy rate following transfer of frozen-thawed embryos were evaluated. Also calving state was investigated. The results obtained were as follows ; The mean number of total and transferrable embryos recovered per superovulated cow was 8.72 and 4.90, respectively, from a total of 72 superovulations using 34 donor cows. There were no significant differences in the number of total or transferrable embryos recovered per superovulated cow between products of follicle stimulating hormone (FSH), years, seasons, and collection numbers. The pregnancy rate was found 44.44% following transfer of frozen-thawed embryos of Korean Native cattle to a total of 180 recipient cows including 82 Angus, 27 Charolais, 62 Hereford and 9 Korean Native cows. The pregnancy rate was significantly (P<0.05) higher in the transfer of excellent (42.99) and good embryos (40.17%), compared with fair (5.90%) grade embryos. And the pregnancy rate was significantly (P<0.05) higher in the transfer of embryos of morula stage (43.86%) than blastocyst stage (15.51%). But there were no significant differences in pregnancy rates between natural and induced estrus estrus asynchrony of 1 days, breeds, and parities of recipient cows. The normal calving rate of 80 pregnant cows following transfer of frozen4hawed em-bryos was 87.5% and the other 10 pregnant cows showed abortion during the period from pregnancy diagnosis at 50~60 days to calving. The average gestation length of normally delivered recipients was 288.50 days and the average birth weight of 70 calves born was 24.22 kg. The gestation length was significantly (P<0.05) shorter in the recipients delivering female calves (286.70 days) than males (289.39 days). But there were no significant differences in gestation tength and birth weight of calves born between the recipient breeds.

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A Study on Properties of CuInS2 Thin Films by Cu/ln Ratio (Cu/In 비에 따른 CuInS2 박막의 특성에 관한 연구)

  • Yang, Hyeon-Hun;Park, Gye-Choon
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.20 no.7
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    • pp.594-599
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    • 2007
  • [ $CulnS_2$ ] thin films were synthesized by sulfurization of Cu/In Stacked elemental layer deposited onto glass Substrates by vacuum furnace annealing at temperature $200^{\circ}C$. And structural and electrical properties were measured in order to certify optimum conditions for growth of the ternary compound semiconductor $CuInS_2$ thin films with non-stoichiometry composition. $CuInS_2$ thin film was well made at the annealed $200^{\circ}C$ of SLG/Cu/In/S stacked elemental layer which was prepared by thermal evaporator, and chemical composition of the thin film was analyzed nearly as the proportion of 1 : 1 : 2. Physical properties of the thin film were investigated at various fabrication conditions substrate temperature, annealing and temperature, annealing time by XRD, FE-SEM and Hall measurement system. The compositional deviations from the ideal chemical formula for $200^{\circ}C$ material can be conveniently described by non-molecularity$({\Delta}x=[Cu/In]-1)$ and non-stoichiometry $({\Delta}y=[{2S/(Cu+3In)}-1])$. The variation of ${\Delta}x$ would lead to the formation of equal number of donor and accepters and the films would behave like a compensated material. The ${\Delta}y$ parameter is related to the electronic defects and would determine the type of the majority charge carriers. Films with ${\Delta}y>0$ would behave as p-type material while ${\Delta}y<0$ would show n-type conductivity. At the sane time, carrier concentration, hall mobility and resistivity of the thin films was $9.10568{\times}10^{17}cm^{-3},\;312.502cm^2/V{\cdot}s\;and\;2.36{\times}10^{-2}\;{\Omega}{\cdot}cm$, respectively.

In vitro Conidial Germination and Mycelial Growth of Fusarium oxysporum f. sp. fragariae Coordinated by Hydrogen Peroxideand Nitric Oxide-signalling

  • Do, Yu Jin;Kim, Do Hyeon;Jo, Myung Sung;Kang, Dong Gi;Lee, Sang Woo;Kim, Jin-Won;Hong, Jeum Kyu
    • The Korean Journal of Mycology
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    • v.47 no.3
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    • pp.219-232
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    • 2019
  • Chemicals related to hydrogen peroxide ($H_2O_2$) and nitric oxide (NO) generations were exogenously applied to Fusarium oxysporum f. sp. fragariae (Fof) causing Fusarium wilt disease in strawberry plants, and regulations of in vitro conidial germination and mycelial growth of the fungus by the chemical treatments were evaluated. $H_2O_2$ drastically reduced the conidial germination of Fof in a dose-dependent manner, and treatment with 3-amino-1,2,4-triazole (3-AT) catalase inhibitor also led to dose-dependent inhibition of conidial germination but relatively moderately. Gradual decreases in mycelial growth of Fof were found by high concentrations of $H_2O_2$, whilst exogenous 3-AT slightly increased the mycelial growth. Increasing sodium nitroprusside (SNP) NO donor, $N^G$-nitro-l-arginine methyl ester (L-NAME) NO synthase (NOS)-inhibitor and tungstate nitrate reductase (NR) inhibitor led to dose-dependent reductions in conidial germination of Fof in quite different levels. SNP conversely increased the mycelial growth but increasing L-NAME moderately decreased the mycelial growth. Tungstate strongly enhanced mycelial growth. Differentially regulated in vitro mycelial growths of Fof were demonstrated by SNP, L-NAME and tungstate with or without $H_2O_2$ supplement. Superoxide anion production was also regulated during the mycelial growth of Fof by nitric oxide. These results show that $H_2O_2$ and NO-associated enzymes can be suggested as fungal growth regulators of Fof as well as eco-friendly disease-managing agents in strawberry production fields.

PKC Downstream of PI3-Kinase Regulates Peroxynitrite Formation for Nrf2-Mediated GSTA2 Induction

  • Kim, Sang-Geon;Kim, Sun-Ok
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.757-762
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    • 2004
  • The protective adaptive response to electrophiles and reactive oxygen species is mediated by the induction of phase II detoxifying genes including glutathione S-transferases (GSTs). NF-E2-related factor-2 (Nrf2) phosphorylation by protein kinase C (PKC) is a critical event for its nuclear translocation in response to oxidative stress. Previously, we have shown that peroxynitrite plays a role in activation of Nrf2 and Nrf2 binding to the antioxidant response element (ARE) via the pathway of phosphatidylinositol 3-kinase (PI3-kinase) and that nitric oxide synthase in hepatocytes is required for GSTA2 induction. In view of the importance of PKC and Pl3-kinase in Nrf2-mediated GST induction, we investigated the role of these kinases in peroxynitrite formation for GSTA2 induction by oxidative stress and determined the relationship between PKC and PI3-kinase. Although PKC activation by phorbol 12-myristate-13-acetate (PMA) did not increase the extents of constitutive and inducible GSTA2 expression, either PKC depletion by PMA or PKC inhibition by staurosporine significantly inhibited GSTA2 induction by tert-butylhydroquinone (t-SHa) a prooxidant chemical. Therefore, the basal PKC activity is req- uisite for GSTA2 induction. 3-Morpholinosydnonimine (SIN-1), which decomposes and yields peroxynitrite, induced GSTA2, which was not inhibited by PKC depletion, but slightly enhanced by PKC activation, suggesting that PKC promotes peroxynitrite formation for Nrf2-mediated GSTA2 induction. Treatment of cells with S-nitroso-N-acetyl-penicillamine (SNAP), an exogenous NO donor, in combination with t-BHQ may produce peroxynitrite. GSTA2 induction by SNAP + t-BHQ was not decreased by PKC depletion, but rather enhanced by PKC activation, showing that the activity of PKC might be required for peroxynitrite formation. LY294002 a P13-kinase inhibitor blocked GSTA2 induction by t-BHQ, which was reversed by PMA-induced PKC activation. These results provide evidence that PKC may playa role in formation of peroxynitrite that activates Nrf2 for GSTA2 induction and that PKC may serve an activator for GSTA2 induction downstream of PI3-kinase.

Hydrogen sulfide restores cardioprotective effects of remote ischemic preconditioning in aged rats via HIF-1α/Nrf2 signaling pathway

  • Wang, Haixia;Shi, Xin;Cheng, Longlong;Han, Jie;Mu, Jianjun
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.3
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    • pp.239-249
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    • 2021
  • The present study explored the therapeutic potential of hydrogen sulfide (H2S) in restoring aging-induced loss of cardioprotective effect of remote ischemic preconditioning (RIPC) along with the involvement of signaling pathways. The left hind limb was subjected to four short cycles of ischemia and reperfusion (IR) in young and aged male rats to induce RIPC. The hearts were subjected to IR injury on the Langendorff apparatus after 24 h of RIPC. The measurement of lactate dehydrogenase, creatine kinase and cardiac troponin served to assess the myocardial injury. The levels of H2S, cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), nuclear factor erythroid 2-related factor 2 (Nrf2), and hypoxia-inducible factor (HIF-1α) were also measured. There was a decrease in cardioprotection in RIPC-subjected old rats in comparison to young rats along with a reduction in the myocardial levels of H2S, CBS, CSE, HIF-1α, and nuclear: cytoplasmic Nrf2 ratio. Supplementation with sodium hydrogen sulfide (NaHS, an H2S donor) and l-cysteine (H2S precursor) restored the cardioprotective actions of RIPC in old hearts. It increased the levels of H2S, HIF-1α, and Nrf2 ratio without affecting CBS and CSE. YC-1 (HIF-1α antagonist) abolished the effects of NaHS and l-cysteine in RIPC-subjected old rats by decreasing the Nrf2 ratio and HIF-1α levels, without altering H2S. The late phase of cardioprotection of RIPC involves an increase in the activity of H2S biosynthetic enzymes, which increases the levels of H2S to upregulate HIF-1α and Nrf2. H2S has the potential to restore aging-induced loss of cardioprotective effects of RIPC by upregulating HIF-1α/Nrf2 signaling.

Reconstruction of a Circumferential Penile Shaft Defect Using Bilateral Superficial External Pudendal Artery Perforator Flaps (양측 천외음부동맥 천공지 피판을 이용한 음경둘레 연조직 결손의 재건)

  • Park, Jongohk;Shin, Donghyeok;Choi, Hyungon;Kim, Jeenam;Lee, Myungchul
    • Archives of Hand and Microsurgery
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    • v.23 no.4
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    • pp.296-300
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    • 2018
  • Penile shaft reconstruction requires adequate soft tissue characteristics as well as constant vascular pedicles when considering a perforator flap. The free flap technique using various donor sites and regional conventional and perforator flaps have been utilized for penile shaft reconstruction. Still, the free flap techniques include challenging surgical procedures in addition to postoperative management. The regional flap can be applied to limited defects due to the size and shape. We performed the bilateral superficial external pudendal artery (SEPA) perforator flaps in order to reconstruct a circumferential penile shaft defect. The circumferential wound has noted necrotic tissue involving superficial (Dartos) fascia. We underwent debridement, preserving deep (Buck's) fascia and corpus spongiosum. Thereafter, the soft tissue defect was covered with bilateral SEPA perforator flaps. The patient has been observed for 27 months, showing penile resilience without deformity or wound-related problems.

Initiating Liver Transplantation at a Public Hospital in Korea

  • Lee, Doo-ho;Lee, Hae Won;Ahn, Young Joon;Kim, Hyeyoung;Yi, Nam-Joon;Lee, Kwang-Woong;Suh, Kyung-Suk
    • Korean Journal of Transplantation
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    • v.31 no.4
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    • pp.193-199
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    • 2017
  • Background: Although there more than 1,000 liver transplantations (LTs) are performed in Korea annually, their immense cost remains a great hurdle. Hence, in an attempt to reduce the medical costs of LT, a program was initiated at a public hospital affiliated with the Seoul National University Hospital. Methods: A total of 11 LTs have been successfully executed since the first LT performed at Seoul Metropolitan Government Seoul National University Boramae Medical Center in July 2011 through December 2014. Results: Nine patients (81.8%) were male and two (18.2%) were female. The mean age of patients was 53.4±11.4 years. Hepatitis B virus-related liver disease (n=6, 54.5%) was the most common causative disease, followed by alcoholic liver disease (ALD) (n=4, 36.4%). The actuarial 3-year survival rate was 90.9%. The median total medical cost of LTs was US $41,583 (calculated from operation to discharge), but only $11,860 was actually charged for patients with health insurance coverage. One female patient who had undergone deceased donor LT for alcoholic liver cirrhosis died during follow-up. This patient was non-compliant with the medical instructions after discharge, and finally expired due to septic shock at 10 months post-LT. Conclusions: In the public hospital, LT was successfully performed at a much lower cost. However, LT guidelines and peritransplant management protocols for patients with ALD must be established before escalating LT at public hospitals since ALD with poor compliance is one of the most common causes of complications at public hospitals.

Glue Embolization of Lymphopseudoaneurysm for Chylous Ascites after Retroperitoneal Surgery

  • Lyo Min Kwon;Saebeom Hur;Chang Wook Jeong;Hwan Jun Jae;Jin Wook Chung
    • Korean Journal of Radiology
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    • v.22 no.3
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    • pp.376-383
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    • 2021
  • Objective: To assess the safety and efficacy of lymphopseudoaneurysm (LPA) glue (n-butyl cyanoacrylate [NBCA]) embolization in the management of chylous ascites after retroperitoneal surgery. Materials and Methods: A retrospective analysis from January 2014 to October 2018 was performed in six patients (4 females and 2 males; mean age, 45.3 ± 14.2 years; range, 26-61 years) who underwent LPA embolization for chylous ascites developing after retroperitoneal surgery involving the perirenal space (four donor nephrectomies, one partial nephrectomy, and one retroperitoneal lymphadenectomy). After placing a percutaneous drainage catheter into the LPA or adjacent lymphocele, embolization was performed by filling the LPA itself with a mixture of glue and Lipiodol (Guerbet). Results: Daily drainage from percutaneously placed drains exceeded 300 mL/day despite medical and surgical treatment (volume: mean, 1173 ± 1098 mL; range, 305-2800 mL). Intranodal lymphangiography was performed in four of the six patients and revealed leakage in 2 patients. Percutaneous embolization of the LPA was successful in all patients using an NBCA and Lipiodol mixture in a ratio of 1:1-1:2 (volume: mean, 4.3 ± 1.1 mL; range, 3-6 mL). Chylous ascites was resolved and the drainage catheter was removed in all patients within 4 days after the procedure (mean, 2.0 ± 1.8 days; range, 0-4 days). No procedure-related complications or recurrence of chylous ascites occurred during a mean follow-up period of 37.3 months (range, 21.1-48.4 months). Conclusion: Glue embolization of LPA has the potential to be a feasible and effective treatment method for the management of chylous ascites after retroperitoneal surgery.