Production and release of pigments by plant hairy root cultures

Taya, Masahito,Tone, Setsuji 한국농화학회 1992 Applied Biological Chemistry (Appl Biol Chem) Vol.35 No.5

Recently, plant hairy roots have become of interest because of their active proliferation in a phytohormonefree medium and their capacity to produce valuable metabolites accumulated at comparable levels of the original plant roots. The purpose of our work is to extend the biotechnological application of hairy root cultures through studies on the following subjects. 1) Induction and characterization of hairy roots: Some hairy root clones were induced from red beet (Beta vulgaris), madder (Rubia tinctorum) and horseradish (Armoraeia lapathifolia) by Agrobacterium rhizogenes infection. These hairy roots had high contents of pigments or enzymes. 2) Bioreactor configuration: A turbine-blade fermentor was developed as a bioreactor suitable for the submerged culture of hiary root with a highly branched morphology. In this fermentor, a stainless steel mesh separated the culture space from the agitation space so that the hairy root cells were not in direct contact with the impellers. 3) Extracellular production of pigments: In the cultures of red beet and madder hairy roots, significant amounts of pigments were released from the cells into medium when the cells were subjected to the treatment of O₂ starvation. Both hairy roots were capable of growing even after the treatment. 4) Morphological change of hairy root: By cultivating the horseradish hairy root in the presence of phytohormones, the cell morphology could be changed reversibly between dedifferentiated-cell aggregates and organized hairy root.

High Production of Recombinant Norwegian Salmonid Alphavirus E1 and E2 Proteins in Escherichia coli by Fusion to Secretion Signal Sequences and Removal of Hydrophobic Domains

Anne Tondervik,Simone Balzer,Tone Haugen,Håvard Sletta,Marit Rode,Karine Lindmo,Trond E. Ellingsen,Trygve Brautaset 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.4

The Norwegian salmonid alphavirus (NSAV)infects farmed Atlantic salmon and rainbow trout, causes pancreas disease and leads to economic losses and fish health issues for the aquaculture. Vaccines are available,but recurring infection outbreaks at Norwegian fish farms have led to endeavours in finding solutions for increased prevention. The NSAV E1 and E2 envelope proteins are potential targets for production of recombinant subunit vaccines and for generation of antibodies for diagnostics. Efficient expression of target proteins is necessary for these applications, and here we present a new strategy for expressing this kinds of viral proteins. We show that 5'-terminal fusion of signal sequences OmpA and CSP to the e1 and e2 genes and removal of the C-terminal hydrophobic interaction and transmembrane domains of E1 and E2 leads to significantly increased expression levels. Recombinant Escherichia coli strains for high-level production of E1 and E2 harbouring these modifications were established using the inducible XylS/Pm expression cassette. Furthermore,reduction of temperature to 16oC after induction leads to 4-fold increase in production for E1, and under high-celldensity cultivations we obtained production levels up to 2.3 g/L. We also show that these proteins can be purified from inclusion bodies by affinity chromatography. This demonstrates the present approach as promising for large scale production of such viral proteins.

Production and Release of Pigments by Plant Hairy Root Cultures

Masahito Taya,Setsuji Tone 한국응용생명화학회 1992 한국응용생명화학회 학술발표회 Vol.1992 No.-

Recently, plant hairy roots have become of interest because of their active proliferation in a phytohormonefree medium and their capacity to produce valuable metabolites accumulated at comparable levels of the original plant roots. The purpose of our work is to extend the biotechnological application of hairy root cultures through studies on the following subjects. 1) Induction and characterization of hairy roots: Some hairy root clones were induced from red beet (Beta vulgaris), madder (Rubia tinctorum) and horseradish (Armoraeia lapathifolia) by Agrobacterium rhizogenes infection. These hairy roots had high contents of pigments or enzymes. 2) Bioreactor configuration: A turbine-blade fermentor was developed as a bioreactor suitable for the submerged culture of hiary root with a highly branched morphology. In this fermentor, a stainless steel mesh separated the culture space from the agitation space so that the hairy root cells were not in direct contact with the impellers. 3) Extracellular production of pigments: In the cultures of red beet and madder hairy roots, significant amounts of pigments were released from the cells into medium when the cells were subjected to the treatment of O₂ starvation. Both hairy roots were capable of growing even after the treatment. 4) Morphological change of hairy root: By cultivating the horseradish hairy root in the presence of phytohormones, the cell morphology could be changed reversibly between dedifferentiated-cell aggregates and organized hairy root.

Quantitative computed tomography measures of skeletal muscle mass in patients with idiopathic pulmonary fibrosis according to the multidisciplinary discussion diagnosis

( Nobuyasu Awano ),( Minoru Inomata ),( Naoyuki Kuse ),( Mari Tone ),( Hanako Yoshimura ),( Tatsunori Jo ),( Kohei Takada ),( Chikatoshi Sugimoto ),( Tomonori Tanaka ),( Hiromitsu Sumikawa ),( Yuzo Su 대한결핵 및 호흡기학회 2019 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.127 No.-

Background: Idiopathic pulmonary fibrosis (IPF) is a progressive disease often accompanied by skeletal muscle wasting. We aimed to assess whether skeletal muscle mass and muscle attenuation on computed tomography (CT) are predictors of mortality in IPF patients using a nationwide cloud-based database and web-based multidisciplinary discussion (MDD) system. Methods: IPF patients diagnosed using MDD from April 2009 to March 2014 were included. We analyzed the cross-sectional area (CSA) of the erector spinae muscle (ESM_CSA) and the pectoralis muscle (PM_CSA), muscle attenuation of ESM (ESM_MA), and PM (PM_MA) on single-slice axial CT. Survival probability was assessed by the Kaplan-Meier method and compared by the log-rank test. Multivariate Cox proportional hazards models were performed to evaluate the relationship among the ESM_CSA, PM_CSA, ESM_MA, PM_MA, clinical parameters, and prognosis. Results: A total of 199 IPF patients were enrolled. Seventy-four patients died during the study period and the most frequent cause was acute exacerbation (13.1%). The group with the lowest quartile of ESMCSA had significantly worse survival than other groups (P = 0.009). Survival rates of the groups with the lowest quartile of PM_CSA, lower ESM_MA, and lower PM_MA were not different from other groups. After multivariate analysis, ESM_CSA < lower quartile was significantly associated with all-cause mortality (hazards ratio, 1.96; P = 0.030), whereas, ESM_MA < median, PM_CSA < lower quartile, and PM_MA < median were not. Conclusions: Low ESM_CSA on CT images may be a strong risk factor for all-cause mortality in IPF patients nationwide by MDD diagnosis.

Design and Stability Analysis of Input Filter for Power Factor Correction Circuits Empolying Discontinuous-Mode Boost Converters

Hsiao Yao Lee,Fu Yuan Shih,Yie Tone Chen,Yan Pei Wu 대한전자공학회 1994 JTC-CSCC : Joint Technical Conference on Circuits Vol.- No.-

ELID, Micro and UltraFabrication Technologies

Hitoshi OHMORI,Y. Hachisu,N. Tone,Y. Uehara,M. Mizutani,S. Wada,T. Ogawa,Y. Takizawa,J. Komotori 한국기계가공학회 2012 한국기계가공학회 춘추계학술대회 논문집 Vol.2012 No.6

황산환원균 Desulfovibrio desulfricans의 혐기배양에 의한 가용성 우라늄의 침전

조규성,주현규,홍고야쓰히로,다야마사히또,도네세쯔지 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.3

황산화원균 D. desulfricans의 배양에 의하여 가용성 우라늄의 침전회수 실험을 진행한 결과를 요약하면 다음가 같다. 먼저 본실험을 위한 균주선정에서 3종의 황산환원균 D. desulfricans, D. multivorans, D. variabilis를 배양, 일정시간 후 탄소원인 젖산(lactate)과 포름산(formate)의 분해속도 및 철이온의 침전량을 조사한 결과 D. desulfricans가 가장 좋았다. 이때 이 균주는 포름산보다 젖산쪽의 이용이 양호하였다. D. desulfricans는 우라늄이 함유된 배양기(60×10exp(-3) ㎏/m^3-broth 이하)에서도 증식에 저해작용이 거의 없었으나, 우라늄이 함유되지 않은 배지에서 증식이 보다 양호하였다. 황산환원균 D. desulfricans는 젖산을 잘 이용(배양 30시간 이내에서의 분해속도 390 mmol/㎥·h)하였으며, pH는 균체 증식과 더불어 pH 9.0까지 증가하였다. 균체수는 배양 70시간에서 6.1×10^14 cells/㎥까지 증식하였다. 균체증식과 더불어서 SO^(2-)_4이온은 환원(8.5mmol/㎥)되어 S^(2-)이온의 생성(7.5 mmol/㎥)이 진행되었다. 배양액 중의 가용성 우라늄 이온은, 황산환원균 D. desulfricans의 무접종구에서는 70시간 이상이 경과하여도 용액중에 그대로 용해되어 있었다. 그러나 균체를 접종배양하였을 경우는 1시간 이내에 용액중의 우라늄은 많은 량이 침전하였다가, 균체의 증식과 더불어 다량의 CO_3^(2-)이온의 생성으로 침전하였던 우라늄의 일부가 용해하였다가, H_2S와 CO_3^(2-) 이온의 축출로 인하여 우라늄 이온은 다시 침전이 되었다. 침전한 우라늄 이온은 70% 이상 대부분이 우라늄 U(VI)로 구성되어 있었다. 따라서 황산환원균 D. desulfricans의 배양에 의하여 가용성 우라늄은, 1) 불용성 Uranyl hydroxide[UO_2(OH)_2] 형태로의 침전, 2) 불용성 Uraninite(UO_2) 형태로 S^(2-)이온 등에 의한 U(VI)에서 U(IV)에로 화원되어 침전, 3) 가용상태의 우라늄 이온이 균체세포벽에 UO_2^(2+)[U(VI)] 형태로 키일레이션 혹은 흡착이 진행되어 침전이 가능하다. The aims of the present study are to examine the precipitation of uranyl ion in the culture of Desulfovibrio desulfricans for the sedimentary recovery of aqueous uranium. D. desulfricans had the highest utilization rate of lactate and precipitated iron ion in the three sulfate reducing bacteria. So, subsequent experiments were conducted using lactate as an energy source. The normal growth was observed with increased pH and lactate utilization. During the culture, the amounts of SO_4^(2-) consumed and S^(2-) produced in aqueous phase were 8.5 and 7.5 mmol/㎥-broth, respectively. More than 85% of soluble uranium precipitated within 1 hour after the inoculation, whereas no uranium precipitation was observed for 70 hours in the culture without inoculum. It was found that the small amount of precipitated uranium was obtained in the alkaline medium with the large amount of Na_2CO_3. Uranyl hydroxide becomes soluble in the presence of CO_3^(2-) with the formation of complex ions such as [UO_2(CO_3)_2·2H_2O]^(2-). Precipitated uranium, both U(VI) and U(IV) were detected, and the former was the main form. In the culture of D. desulfricans soluble uranium (UO_2^(2-)) precipitated in the forms of U(VI) and U(IV). This uranium sedimentation was considered as attributed to the UO_2(OH)_2 and UO_2 formation, and the UO_2^(2+) adsorption to the bacterial cell walls.

Cytostatic in vitro Effects of DTCM-Glutarimide on Bladder Carcinoma Cells

Brassesco, Maria S.,Pezuk, Julia A.,Morales, Andressa G.,De Oliveira, Jaqueline C.,Valera, Elvis T.,Da Silva, Glenda N.,De Oliveira, Harley F.,Scrideli, Carlos A.,Umezawa, Kazuo,Tone, Luiz G. Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.5

Bladder cancer is a common malignancy worldwide. Despite the increased use of cisplatin-based combination therapy, the outcomes for patients with advanced disease remain poor. Recently, altered activation of the PI3K/Akt/mTOR pathway has been associated with reduced patient survival and advanced stage of bladder cancer, making its upstream or downstream components attractive targets for therapeutic intervention. In the present study, we showed that treatment with DTCM-glutaramide, a piperidine that targets PDK1, results in reduced proliferation, diminished cell migration and G1 arrest in 5637 and T24 bladder carcinoma cells. Conversely, no apoptosis, necrosis or autophagy were detected after treatment, suggesting that reduced cell numbers in vitro are a result of diminished proliferation rather than cell death. Furthermore previous exposure to 10 ${\mu}g/ml$ DTCM-glutarimide sensitized both cell lines to ionizing radiation. Although more studies are needed to corroborate our findings, our results indicate that PDK1 may be useful as a therapeutic target to prevent progression and abnormal tissue dissemination of urothelial carcinomas.