• Title/Summary/Keyword: $I_c$ degradation

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Purification and Biological Characterization of Wild-type and Mutants of a Levan Fructotransferase from Microbacterium sp. AL-210 (Microbacterium sp. A-210이 생성하는 Levan fructotransferase의 정제 및 생물학적 특성에 관한 연구)

  • Hwang, Eun-Young;Jeong, Mi-Suk;Cha, Jae-Ho;Jang, Se-Bok
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1218-1225
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    • 2009
  • Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

Inhibitory Effect of Protaetiamycine 9 Derived from Protaetia brevitarsis seulensis Larvae on LPS-mediated Inflammation in RAW264.7 Cells (LPS로 자극한 RAW264.7 대식세포에서 흰점박이꽃무지 유충 유래 Protaetiamycine 9의 항염증 효과)

  • Choi, Ra-Yeong;Seo, Minchul;Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Hwang, Jae-Sam
    • Journal of Life Science
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    • v.31 no.11
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    • pp.987-994
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    • 2021
  • Our previous studies have reported that antimicrobial peptides (AMPs) derived from the larvae of white-spotted flower chafer (Protaetia brevitarsis seulensis) exert anti-inflammatory and neuroprotective activities. This study explored the anti-inflammatory effects of protaetiamycine 9 (CVLKKAYFLTNLKLRG-NH2), a novel AMP, derived from P. b. seulensis against lipopolysaccharide (LPS)-mediated inflammatory response in RAW264.7 macrophage cells. Protaetiamycine 9 (25, 50, 75, and 100 ㎍/ml) did not cause cytotoxic effects against RAW264.7 cells. The RAW264.7 cells were pre-treated with various concentrations of protaetiamycine 9 (25-100 ㎍/ml) for 1 hr and then exposed to LPS (100 ng/ml) for 24 hr. Protaetiamycine 9 treatments decreased the LPS-induced secretion of inflammatory mediators, such as nitric oxide (NO), in a dose-dependent manner. Protaetiamycine 9 (25-100 ㎍/ml) effectively downregulated the LPS-induced increase in mRNA and the protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), which are involved in the production of inflammatory mediators. Protaetiamycine 9 also suppressed the production and gene expression of pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, compared to the presence of LPS alone. Furthermore, protaetiamycine 9 inhibited the degradation of inhibitory kappa B alpha (IκB-α) and the phosphorylation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. In conclusion, these results suggest that protaetiamycine 9 exhibits LPS-mediated inflammatory responses by blocking IκB-α degradation and MAPK phosphorylation.

Influence of Hydrothermal Treatment of Wheat Bran on Phytate-P Content and Performance of Broiler Chickens (수침처리가 밀기울의 피틴태 인 함량과 육계의 생산성에 미치는 영향)

  • Kim, B. H.;Paik, I. K.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.229-240
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    • 2003
  • An in vitro test and a broiler feeding trial were conducted to test the effect of hydrothermal treatment of wheat bran on phytate-P degradation and it’s feeding effect on performance of broilers. Hydrothermal treatment of wheat bran was carried out at 55$^{\circ}C$ with pH 5.5 buffer solution. Phytate-P content of wheat bran decreased quadrically as the wheat bran: buffer solution ratio increased from 1:0.5 to 1:5. Phytate-P degradation was not significantly affected by incubation times above 10 min., drying temperature (55$^{\circ}C$, 65$^{\circ}C$ and 75$^{\circ}C$) or pH of the buffer solution (5.5 and 7.0). A feeding trial was conducted with 240 sex separated d-old broiler chickens (Ross$^{\circledR}$). Broilers were randomly housed to 24 cages of 10 birds each. Six cages (3 of each sex) were assigned to 4 treatments: Control-normal level of non-phytate-P (NPP); LP-low NPP treatment which had 0.1% lower NPP than Control; LPWB-LP with wheat bran which provided 475 IU of plant phytase per kg diet; LPHWB-LP with hydrothermally treated wheat bran. Results of the feeding trial showed that broilers in the LP treatment gained significantly less than other treatments in starter period (1-21d) but only male broilers for growing LP gained significantly less than Control in the grower (22-35d) and overall period. There were no significant differences in weight gain among the birds of LPWB, LPHWB and Control. Feed intake during the overall period was not significantly different between LPWB and Control but that of LP was lower than LPHWB and that of LPHWB was lower than Control. Feed/gain ratio was significantly lower in LPHWB and LP than in Control and LPWP. Mortality was highest in LPHWB. Availability of crude fat, crude ash and Ca was significantly lower in LP than other treatments. Availability of P and Zn was higher in LPWB and LPHWB than in Control and LP. Availability of P, Mg and Zn was highest in LPHWP treatment. Excretion of P was significantly lower in low NPP treatments than in Control. Serum Ca level was highest whereas serum P level was lowest in LP. Tibial crude ash content was higher in wheat bran treatments, but lower in LP than Control. However, tibial Ca content was higher in Control and LP than wheat bran treatments. Tibial P content of LP and LPWB was lower than Control. However, tibial content of Fe was highest in LP. It was concluded that wheat bran, a source of plant phytase, could be used in low NPP broiler diets to prevent the depression of performance. Reduction of P excretion can be achieved concomitantly. Hydrothermal treatment of wheat bran was effective in improving utilizability of some minerals but was not effective in improving performance of broilers.

Plumbagin from Plumbago Zeylanica L Induces Apoptosis in Human Non-small Cell Lung Cancer Cell Lines through NF-κB Inactivation

  • Xu, Tong-Peng;Shen, Hua;Liu, Ling-Xiang;Shu, Yong-Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2325-2331
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    • 2013
  • Objective: To detect effects of plumbagin on proliferation and apoptosis in non-small cell lung cancer cell lines, and investigate the underlying mechanisms. Materials and Methods: Human non-small cell lung cancer cell lines A549, H292 and H460 were treated with various concentrations of plumbagin. Cell proliferation rates was determined using both cell counting kit-8 (CCK-8) and clonogenic assays. Apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry and TUNEL assay. The levels of reactive oxygen species (ROS) were detected by flow cytometry. Activity of NF-${\kappa}B$ was examined by electrophoretic mobility shift assay (EMSA) and luciferase reporter assay. Western blotting was used to assess the expression of both NF-${\kappa}B$ regulated apoptotic-related gene and activation of p65 and $I{\kappa}B{\kappa}$. Results: Plumbagin dose-dependently inhibited proliferation of the lung cancer cells. The IC50 values of plumbagin in A549, H292, and H460 cells were 10.3 ${\mu}mol/L$, 7.3 ${\mu}mol/L$, and 6.1 ${\mu}mol/L$ for 12 hours, respectively. The compound concentration-dependently induced apoptosis of the three cell lines. Treatment with plumbagin increased the intracellular level of ROS, and inhibited the activation of NK-${\kappa}B$. In addition to inhibition of NF-${\kappa}B$/p65 nuclear translocation, the compound also suppressed the degradation of $I{\kappa}B{\kappa}$. ROS scavenger NAC highly reversed the effect of plumbagin on apoptosis and inactivation of NK-${\kappa}B$ in H460 cell line. Treatment with plumbagin also increased the activity of caspase-9 and caspase-3, downregulated the expression of Bcl-2, upregulated the expression of Bax, Bak, and CytC. Conclusions: Plumbagin inhibits cell growth and induces apoptosis in human lung cancer cells through an NF-${\kappa}B$-regulated mitochondrial-mediated pathway, involving activation of ROS.

Change of Antioxidative Activity at Different Harvest Time and Improvement of Atopic Dermatitis Effects for Persimmon Leaf Extract (감잎(柿葉) 추출물의 채취시기에 따른 항산화 활성 변화 및 아토피성 피부염 개선효과)

  • Jung, Woo-Young;Jeong, Jong-Moon
    • The Korea Journal of Herbology
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    • v.27 no.1
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    • pp.41-49
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    • 2012
  • Objectives : The purpose of this study was to investigate the anti-oxidative and anti-atopic dermatitis effects of persimmon leaf extract obtained from Cheongdo-gun, where more than 60% of Korean persimmon is produced. Methods : Anti-oxidative effects of the crude persimmon leaf extract harvested monthly between May and November were determined by in vitro assay using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide dismutase (SOD)-like reaction. Anti-atopic dermatitis effects of the crude persimmon leaf extract were determined by using collagenase type I inhibition assay and by quantitative assays including serum histamine, prostaglandin E metabolite and leukotriene $B_4$ levels in animal model of atopic dermatitis using Balb/c mice. Results : Persimmon leaf extract harvested in May had higher levels of total phenolic compounds (182.24 mg/g) and flavonoids (23.05 mg/g) than the ones of different month extract. Also, persimmon leaf extract harvested in May showed the most effective extract scavenging activities of DPPH free radical ($13.39{\pm}0.21\;{\mu}g/ml$) and superoxide anion radical ($40.52{\pm}2.32\;{\mu}g/ml$), leading to use the persimmon leaf extract harvested in May for the experiments hereafter. Persimmon leaf extract showed $326.71{\pm}4.6\;{\mu}g/ml$ of 50% inhibitory concentration ($IC_{50}$) for collagenase type I which is responsible for the degradation of extracellular matrix. In addition, persimmon leaf extract application group could decrease serum levels of histamine, prostaglandin E metabolite and leukotriene $B_4$ compared to the negative control in animal model of atopic dermatitis. Especially, persimmon leaf extract showed a significantly decreased serum leukotriene $B_4$ level relative to the levels of histamine and prostaglandin E metabolite. Conclusions : Persimmon leaf extract showed anti-oxidative and anti-atopic dermatitis effects in vitro and in vivo. These results suggest that persimmon leaf extract may have immunoregulatory function for alleviating atopic dermatitis by decreasing collagenase activity and mast cell activation.

Physicochemical Characteristics of Silk Fibroin Degummed by Protease in Bacillus licheniformis I. Physicochemical Characteristics of Degummed Silk Fiber (Bacillus licheniformis 단백질 분해 효소에 의한 정연 견사의 특성 I. 정연 견사의 이화학적 특성)

  • 김영대;남중희
    • Journal of Sericultural and Entomological Science
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    • v.34 no.2
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    • pp.41-51
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    • 1992
  • In this thesis, both soap and enzymatic degumming method were adopted and the optimum degumming conditions were obtained. Difference between the two degumming methods in silk fiber state was investigated and analyzed on the basis of the results of physical testings, polarizing microscopy, scanning electron microscopy, viscosity measurement, (${\alpha}$$\varepsilon$) amino group contents measurement, birefringence measurement, amino acid analysis, thermal analysis, infrared spectroscopy and x-ray diffraction analysis. The results obtained were summarized as follows; Physical test results of the degummed silk fiber showed that the tenacity and the elongation of enzymatic degummed silk fiber were lower than those of soap degummed fiber. But SEM observation and amino acid analysis showed almost the same tendency in the two degumming methods. The viscosity of enzymatic degummed silk fiber was lower than that of soap degummed fiber, but (${\alpha}$$\varepsilon$) amino group contents was higher in the enzymatic degummed fiber. It can be suggested that the enzymatic degummed silk fibroin was more degraded than the soap degummed fibroin. The birefringence, endothermic temperature of DSC spectrum, IR crystallinity and X-ray lateral order factor of enzymatic degummed silk fiber were higher than those of soap degummed fiber. It seems that the enzymatic degummed silk fiber has the higher crystallinity than that of soap degummed one according to the above results. However, it can be inferred that these differences between soap and enzymatic degummed fiber would be lessened if pretreatment and aftertreatment were included in the enzymatic degumming process.

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Effects of a Dietary Chitosan-Alginate-Fe(II) Complex on Meat Quality of Pig Longissimus Muscle during Ageing

  • Park, B.Y.;Kim, J.H.;Cho, S.H.;Hwang, I.H.;Jung, O.S.;Kim, Y.K.;Lee, J.M.;Yun, S.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.3
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    • pp.414-419
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    • 2005
  • The current study was conducted to investigate the effects of dietary chitosan-alginate-Fe(II) complex (CAFC) supplementation on carcass and meat qualities of pig m. longissimus during chiller ageing. One hundred and twenty-two LYD (Landrace${\times}$Yorkshire${\times}$Duroc) pigs were sampled from an industrial population. Seventy-four pigs (32 gilts and 42 barrows) were administered 3 ml of dietary supplementation of CAFC per day from 25 to 70 days of age, while the remaining 48 pigs (20 gilts and 28 barrows) were fed the same commercial feeding regime without the supplementation. For assessing the dietary effects on pH, objective meat color, cooking loss, water-holding capacity (WHC), thiobarbituric acid reactive substances (TBARS), volatile basic nitrogen (VBN) and fatty acid composition during ageing, 20 barrows (10 of each treatment) were randomly sampled, and aged for 3, 7, 12, 16, 20 and 25 days in a $1^{\circ}C$ chiller. The results showed that CAFC-fed pigs required approximately 10 fewer feeding days than the control group. Furthermore, the treatment resulted in greatly higher carcass grade whereby the grade A was increased by approximately 35% and 7% for gilts and barrows, respectively. The treatment had no significant effect (p>0.05) on pH, meat color and WHC during ageing. On the other hand, the CAFC-fed pigs showed significantly (p<0.05) lower TBARS values from 20 days of storage. In addition, the sum of unsaturated fatty acids for the treated group was significantly (p<0.05) higher than that for the control group after the storage time. This implied that CAFC supplementation could reduce the formation of free radicals in fatty acids (i.e., lipid oxidation). The treatment also significantly (p<0.05) retarded VBN formation during ageing, indicating a significant reduction in protein degradation. However, as there was no difference in pH between the two groups, the result raised a possibility that antibacterial activity of the CAFC alone could cause reduction in the formation of TBARS and VBN. In this regard, although the treatment effectively slowed down the formation of TBARS and TBA during chiller ageing, it was not resolved whether that was associated with the direct effect of the antioxidant function of chitosan and/or alginate, or a consequence of their antibacterial functions.

Design and Development of VDL Mode-2 D8PSK Modem (VDL Mode-2 D8PSK 모뎀 설계 및 개발)

  • Gim, Jong-Man;Choi, Seoung-Duk;Eun, Chang-Soo
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.34 no.11C
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    • pp.1085-1097
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    • 2009
  • We present a structure and design method of the D8PSK modem compatible with the VDL mode-2 standard and performance test results of the developed modem. In VDL mode-2, the raised cosine filter is used only in the transmitter and a general low pass filter is used in the receiver. Consequently, we can not achieve ISI reduction but can have better spectrum characteristics. Although there is 1~2 dB performance degradation with an un-matched filter compared to that with a matched filter, it is more important to minimize adjacent channel interference in narrow band communications. The transmit signal is generated digitally to avoid the problems(I/Q imbalance and DC offset etc.) of analog modulators. In addition the digital down converter using digital IF sampling technique is adopted for the receiver. This paper contains the overall configuration, design method and simulation results based in part on the previously proposed structures and algorithms. It is confirmed that the modem transmits and receives messages successfully at a speed of max. 870 km/h over ranges of up to 310 km through the ground and in-flight communication tests.

A Comparative Study on Degradation of BTEX Vapor by O3/UV, TiO2/UV, and O3/TiO2/UV System with Operating Conditions (운전조건에 따른 O3/UV, TiO2/UV 및 O3/TiO2/UV 시스템의 BTEX 증기처리에 관한 비교 연구)

  • Kim, Kyoung-Jin;Park, Ok-Hyun
    • Journal of Korean Society for Atmospheric Environment
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    • v.24 no.1
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    • pp.91-99
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    • 2008
  • A multilayer tower-type photoreactor, in which $TiO_2$-coated glass-tubes were installed, was used to measure the vapor-phase BTEX removal efficiencies by ozone oxidation ($O_3$/UV), photocatalytic oxidation ($TiO_2$/UV) and the combination of ozone and photocatalytic oxidation ($O_3/TiO_2$/UV) process, respectively. The experiments were conducted under various relative humidities, temperatures, ozone concentrations, gas flow rates and BTEX concentrations. As a result, the BTEX removal efficiency and the oxidation rate by $O_3/TiO_2$/UV system were highest, compared to $O_3$/UV and $TiO_2$/UV system. The $O_3/TiO_2$/UV system accelerated the low oxidation rate of low-concentration organic compounds and removed organic compounds to a large extent in a fixed volume of reactor in a short time. Therefore, $O_3/TiO_2$/UV system as a superimposed oxidation technology was developed to efficiently and economically treat refractory VOCs. Also, this study demonstrated feasibility of a technology to scale up a photoreactor from lab-scale to pilot-scale, which uses (i) a separated light-source chamber and a light distribution system, (ii) catalyst fixing to glass-tube media, and (iii) unit connection in series and/or parallel. The experimental results from $O_3/TiO_2$/UV system showed that (i) the highest BTEX removal efficiencies were obtained under relative humidity ranging from 50 to 55% and temperature ranging from 40 to $50^{\circ}C$, and (ii) the removal efficiencies linearly increased with ozone dosage and decreased with gas flow rate. When applying Langmuir-Hinshelwood model to $TiO_2$/UV and $O_3/TiO_2$/UV system, reaction rate constant for $O_3/TiO_2$/UV system was larger than that for $TiO_2$/UV system, however, it was found that adsorption constant for $O_3/TiO_2$/UV system was smaller than that for $TiO_2$/UV system due to competitive adsorption between organics and ozone.

Synthesis and Micellar Characterization of CBABC Type PLGA-PEO-PPO-PEO-PLGA Pentablock Copolymers

  • Seong, Haseob;Cho, Eun-Bum;Oh, Joongseok;Chang, Taihyun
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2342-2348
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    • 2014
  • Poly(lactic-co-glycolic acid) (PLGA) were grafted to both ends of Pluronic$^{(R)}$ F68 ($(EO)_{75}(PO)_{30}(EO)_{75}$) triblock copolymer to produce poly{(lactic acid)$_m$-co-(glycolic acid)$_n$}-b-poly(ethylene oxide)$_{75}$-b-poly(propylene oxide)$_{30}$-b-poly(ethylene oxide)$_{75}$-b-poly{(lactic acid)$_m$-co-(glycolic acid)$_n$} (PLGA-F68-PLGA) pentablock copolymers. Molecular weights of PLGA blocks were controlled and five kinds of pentablock copolymers with different PLGA block lengths were synthesized using in-situ ring-opening polymerization of D,L-lactide and glycolide with tin(II) 2-ethylhexanoate ($Sn(Oct)_2$) catalyst. PLGA-F68-PLGA pentablock copolymers were characterized by $^1H$- and $^{13}C$-NMR, GPC, and TGA. The numbers (2m, 2n) of repeating units for lactic acid and glycolic acid inside PLGA segments were obtained as (48, 17), (90, 23), (125, 40), (180, 59), and (246, 64), with $^1H$-NMR measurement. From NMR data, the resultant molecular weights were determined in the range of 12,700-29,700, which were similar to those obtained from GPC. Polydispersity index was increased in the range of 1.32-1.91 as the content of PLGA blocks increased. TG and DTG thermograms showed discrete degradation traces for PLGA and F68 blocks, which indicate the weight fractions of PLGA blocks in pentablock copolymers can be calculated by TG profile and it is possible to remove PLGA block selectively. Hydrodynamic radius and radius of gyration of pentablock copolymer micelle were obtained in the range of 46-68 nm and 31-49 nm, respectively, in very dilute (i.e. 0.005 wt %) aqueous solution of THF:$H_2O$ = 10:90 by volume at $25^{\circ}C$.