• Title/Summary/Keyword: Macromolecule

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Addition of Macromolecules to PZM-3 Culture Medium on the Development and Hatching of In vitro Porcine Embryos

  • Kim, J.Y.;Kim, S.B.;Park, M.C.;Park, H.;Park, Y.S.;Park, H.D.;Lee, J.H.;Kim, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.12
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    • pp.1820-1826
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    • 2007
  • In this study, we conducted various experiments in order to develop enhanced cultural conditions for in vitro-produced porcine embryos. All embryos were produced by in vitro maturation (IVM) and fertilization (IVF) of immature oocytes from abattoir-derived ovaries. In experiment 1, we cultured IVF embryos in 4 different groups, namely, 0% bovine serum albumin (BSA), 3% BSA, 0.05% Polyvinyl alcohol (PVA), and 0.5% Polyvinylpyrrolidone (PVP) added to the basal fluid cultural medium, Porcine zygote medium 3 (PZM-3). The rates of embryo development were higher in the group where the PZM-3 media had been supplemented with 3% BSA than the other groups. While not statistically significant, the percent of blastocysts and hatched blastocytes were 6.9% and 25.0% in the 3% BSA group vs. 1.2-6.4% and 0-16.7% in the other groups, respectively. In experiment 2, we added 10% fetal bovine serum (FBS) to PZM-3 on day 0 of culture and observed the development rate of blastocysts per day of culture from days 0 to 5. The development rate of blastocysts was higher at 15.6% on day 4 than on any other day, and was significantly higher than on day 0 or day 1 (p<0.05). The development rate of hatched blastocysts was 26.7% on day 4, and was higher than on any other day. In experiment 3, we cultured IVF embryos with different fluid culture media, grouped as 1) PZM-3+0.3% BSA (day0-day7); 2) PZM-3+0.3% BSA${\rightarrow}$day-4) PZM-3+10% FBS; 3) PZM-3+0.3% BSA${\rightarrow}$PZM-3+0.3% BSA+(day-4) FBS 10%; and 4) PZM-3+0.3% BSA+10% FBS (day0-day7). The development rates of blastocysts and hatched blastocysts were 21.5% and 53.1% in group 3, respectively, which was significantly higher than group 4 with respect to blastocyst development (5.2%, p<0.05) but not hatched blastocysts (14.3%). The total cell number (TCN) of blastocysts in group 3 was higher at $37.8{\pm}16.1$ than the other groups at $16.8{\pm}4.4$ - $30.1{\pm}10.9$; however, this was not significantly different. The results of this study showed that PZM-3 containing 0.3% BSA and supplemented with FBS during the later stage of culture on day 4 resulted in better TCNs and an increased rate of hatched blastocysts.

Recovery of a High Molecular Soluble Protein from Surimi Wastewater Using Calcium Powder of Cuttle Bone (갑오징어갑 칼슘을 이용한 Surimi 가공폐수로부터 단백질의 회수)

  • KIM Jin-Soo;CHO Moon-Lae;HEU Min-Soo;CHOI Yeung Joon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.2
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    • pp.80-87
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    • 2003
  • Recovery conditions and characteristics of a high molecular soluble protein from surimi processing wastewater in marine manufacture using calcium powder of cuttle bone treated with acetic acid (ATC) were examined. Judging from results of total-N, pH, COD, turbidity and yields, optimal treatment concentration of ATC for recovery of high molecular soluble proteins from wastewater was $1.0\%.$ The protein recovered from seafood waste (PRW) was macromolecule weight. The COD value in the wastewater treated with ATC was very high. The PRW had a $78.4\%$ in moisture, $1.0\%$ in crude lipid and $5.7\%$ in crude ash. The proximate composition, except the crude ash, of the PRW was similar to that of commercial surimi. The PRW showed white index and similar in the content and in the composition of total amino acid to those of commercial surimi. From the results of sensory evaluation on white index and texture, the heat-induced surimi gel prepared with $5\%$ subsititution of the PRW for bulking agent of commercial surimi was not significantly different compared to that prepared with the original commercial surimi.

Characteristics of Fluorescent Organic Matter and Amino Acids Composition in the East Sea (동해의 용존유기물 형광특성 및 아미노산 조성에 관한 연구)

  • 박용철;손승규
    • 한국해양학회지
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    • v.30 no.4
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    • pp.341-354
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    • 1995
  • Fluorescence characteristic and amino acids composition of organic matter were determined from extracted seawater samples at eight stations in the East Sea of Korea. Organic compounds have been extracted onto C-18 Sep-Pak cartridges. Three dimensional excitation/emission fluorescence contouring of extracts showed two markedly distinct characterized fluoroscopies representing protein-like biomacromolecule and humic-like geomacromolecule. Protein-like biomacromolecule showing fluorescence maxima at 280 nm/330 nm (excitation/emission) were abundant in the surface mixed layer and then apparently decreased below the thermocline at most stations. It suggests that source of biomacromolecule is comely related with vigorous biological synthetic activity in the surface layer and bacteria decompose its biologically labile components near the thermocline and in the deeper layer. On the other hand, humiliate geomacromolecule showing fluorescence maxima at 330 nm/430 nm (excitation/emission) were low in the surface mixed layer implying photochemical oxidation and then increased below the thermocline at most stations. It suggests that geomacromolecule might be transformed by condensation of bio-refractoryorganic fraction after decomposition of biomacromolecule and particulate organic carbon derived from the surface mixed layer. HPLC measurements of amino acids showed similar composition between seawater and extracted organic macromolecule after hydrolysis. Glycine, serine and alanine were predominant, accounting for more than 50% of total amino acids. Dissolved free amino acids of seawater were more abundant in the surface layer(0.7∼1.8 uM) than the deeper layer (0.2∼0.4 uM). D/L racemic ratio of alanine of extracted organic matter showed lower value in the surface layer than the deeper layer. It suggests that biomacromolecule predominant in the surface layer is relatively young, rapidly recycling and biologically labile.

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The Separation of Particulate within PFC Decontamination Wastewater Generated by PFC Decontamination (PFC 제염 후 발생된 제염폐액 내 오염입자의 제거)

  • Kim Gye-Nam;Lee Sung-Yeol;Won Hui-Jun;Jung Chong-Hun;Oh Won-Zin;Park Jin-Ho;narayan M.
    • Proceedings of the Korean Radioactive Waste Society Conference
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    • 2005.06a
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    • pp.32-39
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    • 2005
  • When PFC(Perfluorocarbonate) decontamination technology is applied to removal of radioactive contaminated particulate adhered at surface during the operation of nuclear research facilities, it is necessary to develop a filtration equipment to reuse of PFC solution due to high price, also to minimize the volume of second wastewater. Contaminated characteristics of hot particulate was investigated and a filtration process was presented to remove suspended radioactive particulate from PFC decontamination wastewater generated on PFC decontamination. The range of size of hot particulate adhered at the surface of research facilities measured by SEM was $0.1{\sim}10{\mu}m$. Hot particulate of more than $2{\mu}m$ in PFC contamination wastewater was removed by first filter and then hot particulate of more than $0.2{\mu}m$ was removed by second filter. Results of filter experiments showed that filtration efficiency of PVDF(Poly vinylidene fluoride), PP(Polypropylene), Ceramic filter was $95{\sim}97\%$. A ceramic filter showed a higher filtration efficiency with a little low permeate volume. Also, a ceramic of inorganic compound could be broken easily on experiment and has a high price but was highly stable at radioactivity in comparison of PVDF and PP of a macromolecule which generate $H_2$ gas in alpha radioactivity environment.

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In vivo Pharmacokinetics, Activation of MAPK Signaling and Induction of Phase II/III Drug Metabolizing Enzymes/Transporters by Cancer Chemopreventive Compound BHA in the Mice

  • Hu, Rong;Shen, Guoxiang;Yerramilli, Usha Rao;Lin, Wen;Xu, Changjiang;Nair, Sujit;Kong, Ah-Ng Tony
    • Archives of Pharmacal Research
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    • v.29 no.10
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    • pp.911-920
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    • 2006
  • Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around $10\;{\mu}M$. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, ${\gamma}-GCS$, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slco1b2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers.

The Mycobiota of Air Inside and Outside the Meju Fermentation Room and the Origin of Meju Fungi

  • Kim, Dae-Ho;Kim, Sun-Hwa;Kwon, Soon-wo;Lee, Jong-Kyu;Hong, Seung-Beom
    • Mycobiology
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    • v.43 no.3
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    • pp.258-265
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    • 2015
  • The fungi on Meju are known to play an important role as degrader of macromolecule of soybeans. In order to elucidate the origin of fungi on traditional Meju, mycobiota of the air both inside and outside traditional Meju fermentation rooms was examined. From 11 samples of air collected from inside and outside of 7 Meju fermentation rooms, 37 genera and 90 species of fungi were identified. In outside air of the fermentation room, Cladosporium sp. and Cladosporium cladosporioides were the dominant species, followed by Cladosporium tenuissimum, Eurotium sp., Phoma sp., Sistotrema brinkmannii, Alternaria sp., Aspergillus fumigatus, Schizophyllum commune, and Penicillium glabrum. In inside air of the fermentation room, Cladosporium sp., Aspergillus oryzae, Penicillium chrysogenum, Asp. nidulans, Aspergillus sp., Cla. cladosporioides, Eurotium sp., Penicillium sp., Cla. tenuissimum, Asp. niger, Eur. herbariorum, Asp. sydowii, and Eur. repens were collected with high frequency. The concentrations of the genera Aspergillus, Eurotium, and Penicillium were significantly higher in inside air than outside air. From this result and those of previous reports, the origin of fungi present on Meju was inferred. Of the dominant fungal species present on Meju, Lichtheimia ramosa, Mucor circinelloides, Mucor racemosus, and Scopulariopsis brevicaulis are thought to be originated from outside air, because these species are not or are rarely isolated from rice straw and soybean; however, they were detected outside air of fermentation room and are species commonly found in indoor environments. However, Asp. oryzae, Pen. polonicum, Eur. repens, Pen. solitum, and Eur. chevalieri, which are frequently found on Meju, are common in rice straw and could be transferred from rice straw to Meju. The fungi grow and produce abundant spores during Meju fermentation, and after the spores accumulate in the air of fermentation room, they could influence mycobiota of Meju fermentation in the following year. This could explain why concentrations of the genera Aspergillus, Eurotium, and Penicillium are much higher inside than outside of the fermentation rooms.

The Synthesis and MR Properties of New Macromolecular MR Contrast Agent (새로운 거대분자 MR 조영제의 합성 및 MR 특성에 관한 연구)

  • 장용민;장영환;황문정;박현정;전경녀;이종민;배경수;강봉석
    • Investigative Magnetic Resonance Imaging
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    • v.6 no.1
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    • pp.35-40
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    • 2002
  • Purpose : To evaluate the NMR relaxation properties and imaging characteristics of tissue-specificity for a newly developed macromolecular MR agent. Materials and methods : Phthalocyanine (PC) was chelated with paramagnetic ion, Mn.2.01g (5.2 mmol) of Phthalocyanine was mixed with 0.37g (1.4 mmol) of Mn chloride at $310^{\circ}C$ for 36 hours and then purified by chromatography (CHC13/CH3OH 98/2 v/v, Rf, 0.76) to obtain 1.04g (46%) of MnPC (molecular weight= 2000d). The $T1}T2$ relaxivity of MnPC was measured in 1.5T(64 MHz) MR using 0.1 mM MnPC. The MR image characteristics of MnPC was evaluated using spin-echo (TR/TE=500/14 msec) and gradient-echo (FLASH) (TR/TE=80/4 msec, flip angle=60) techniques in 1.57 MR scanner. The images of rabbit liver were obtained every 10 minutes up to 4 hours. To study the effect of concentration on image, 20 mM, 50 mM, 100 mM of MnPC were tested. Results : The relaxivities of MnPC at 1.5T(64MHz) were Rl=7.28 $mM^{-1}S^{-1},{\;}R2=55.56mM^{-1}S^{-1}$. Compared to the values of Gd-DTPA (Rl[=4.8 $mM^{-1}S^{-1})$], R2[=5.2 $mM^{-1}S^{-1}])$]), both T1/T2 relaxivities of MnPC were higher than those of Gd-DTPA. For both of SE and FLASH techniques, the contrast enhancement reached maximum at 10 minutes after bolus injection and the enhancement continued for more than 2 hours. When compared with small molecular weight liver agents such as Gd-EOB-DTPA, Gd-BOPTA and MnDPDP, MnPC was characterized by more prolonged enhancement time. The time course of MR images also revealed biliary excretion of MnPC. Conclusion : We developed a new macromolecular MR agent, MnPC. The relaxivities of MnPC were higher than those of small molecular weight Gd-chelate. Hepatic uptake and biliary excretion of MnPC suggests that this agent is a new liver-specific MR agent.

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Structural Characteristics and Anti-inflammatory Activities of Chemically Sulfated-hyaluronic Acid from Streptococcus dysgalactiae (Streptococcus dysgalactiae로부터 분리된 히알루론산과 황화된 유도체의 구조와 항염증 활성)

  • Hong, Chang-Il;Jung, Eui-Gil;Han, Kook-Il;Kim, Yong Hyun;Lee, Sung Hee;Lee, Hong Sub;Han, Man-Deuk
    • Journal of Life Science
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    • v.26 no.5
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    • pp.545-554
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    • 2016
  • Hyaluronic acid (HA) is an important macromolecule in medical and pharmaceutical fields. HA is a natural and linear polymer composed of repeating disaccharide units of β-1, 3-N-acetyl glucosamine and β-1, 4-glucuronic acid. This work aimed to confirm the structural characteristics and anti-inflammatory activities of HA and its chemically sulfated-HA. HA was produced from a fed-batch fermentation process using Streptococcus dysgalactiae in a 5 l bioreactor. HA was isolated water-soluble form (HA-WS) and water-insoluble form (HA-WI) from culture medium, and was obtained chemically sulfated-derivative (S-HA) that resulted in a 90% yield from HA-WI. The structural features of the sulfated- HA (S-HA) were investigated by FT-IR and 1H-NMR spectroscopy. The FT-IR and NMR patterns revealed the similarity in both the FTIR spectrum as well as NMR spectrum of both reference standard and purified HA from S. dysgalactiae. The anti-inflammatory activities of HA and S-HA were examined on LPS-induced RAW 264.7 cells. S-HA was significantly inhibited production of pro-inflammatory mediators such as nitric oxide (NO) and PGE2 and the gene levels of iNOS and COX-2, which are responsible for the production of NO and PGE2, respectively. Furthermore, S-HA also suppressed the overproduction of pro-inflammatory cytokine TNF-α (<80 pg/ml) and IL-6 (<100 pg/ml) compared to that of HA-WI. The present study clearly demonstrates that HA-S exhibits anti-inflammatory activities in RAW 264.7 macrophage cells.

Quantitative Analysis of Magnetization Transfer by Phase Sensitive Method in Knee Disorder (무릎 이상에 대한 자화전이 위상감각에 의한 정량분석법)

  • Yoon, Moon-Hyun;Sung, Mi-Sook;Yin, Chang-Sik;Lee, Heung-Kyu;Choe, Bo-Young
    • Investigative Magnetic Resonance Imaging
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    • v.10 no.2
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    • pp.98-107
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    • 2006
  • Magnetization Transfer (MT) imaging generates contrast dependent on the phenomenon of magnetization exchange between free water proton and restricted proton in macromolecules. In biological materials in knee, MT or cross-relaxation is commonly modeled using two spin pools identified by their different T2 relaxation times. Two models for cross-relaxation emphasize the role of proton chemical exchange between protons of water and exchangeable protons on macromolecules, as well as through dipole-dipole interaction between the water and macromolecule protons. The most essential tool in medical image manipulation is the ability to adjust the contrast and intensity. Thus, it is desirable to adjust the contrast and intensity of an image interactively in the real time. The proton density (PD) and T2-weighted SE MR images allow the depiction of knee structures and can demonstrate defects and gross morphologic changes. The PD- and T2-weighted images also show the cartilage internal pathology due to the more intermediate signal of the knee joint in these sequences. Suppression of fat extends the dynamic range of tissue contrast, removes chemical shift artifacts, and decreases motion-related ghost artifacts. Like fat saturation, phase sensitive methods are also based on the difference in precession frequencies of water and fat. In this study, phase sensitive methods look at the phase difference that is accumulated in time as a result of Larmor frequency differences rather than using this difference directly. Although how MT work was given with clinical evidence that leads to quantitative model for MT in tissues, the mathematical formalism used to describe the MT effect applies to explaining to evaluate knee disorder, such as anterior cruciate ligament (ACL) tear and meniscal tear. Calculation of the effect of the effect of the MT saturation is given in the magnetization transfer ratio (MTR) which is a quantitative measure of the relative decrease in signal intensity due to the MT pulse.

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The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.