• Asian-Australasian Journal of Animal Sciences
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• 제28권4호
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• pp.559-567
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• 2015

Ex vivo digestion of proteins and fat in Red Chittagong Cattle milk from Bangladesh was carried out using human gastrointestinal enzymes. This was done to investigate the protein digestion in this bovine breed's milk with an especial focus on the degradation of the allergenic milk proteins; ${\alpha}_{s1}$-casein and ${\beta}$-lactoglobulin and also to record the generation of peptides. Lipolysis of the milk fat and release of fatty acids were also under consideration. After 40 min of gastric digestion, all the ${\alpha}_s$-caseins were digested completely while ${\beta}$-lactoglobulin remained intact. During 120 min of duodenal digestion ${\beta}$-lactoglobulin was reduced, however, still some intact ${\beta}$-lactoglobulin was observed. The highest number of peptides was identified from ${\beta}$-casein and almost all the peptides from ${\kappa}$-casein and ${\beta}$-lactoglobulin were identified from the gastric and duodenal samples, respectively. No lipolysis was observed in the gastric phase of digestion. After 120 min of duodenal digestion, milk fat showed 48% lipolysis. Medium (C10:0 to C16:0) and long (${\geq}C17:0$) chain fatty acids showed 6% to 19% less lipolysis than the short (C6:0 to C8:0) chain fatty acids. Among the unsaturated fatty acids $C18:1{\sum}others$ showed highest lipolysis (81%) which was more than three times of $C18:2{\sum}all$ and all other unsaturated fatty acids showed lipolysis ranging from 32% to 38%. The overall digestion of Bangladeshi Red Cattle milk was more or less similar to the digestion of Nordic bovine milk (Norwegian Red Cattle).

• 한국식품영양학회지
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• 제31권3호
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• pp.378-387
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• 2018

This study investigated the volatile flavor composition of essential oils from Chrysanthemum zawadskii var. latilobum Kitamura and Aster yomena Makino. The essential oils obtained by the hydrodistillation extraction method from the aerial parts of the plants were analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). One hundred (95.04%) volatile flavor compounds were identified in the essential oil from the C. zawadskii var. latilobum Kitamura. The major compounds were valencene (10.82%), ${\delta}$-cadinol (9.77%), hexadecanoic acid (8.70%), 2-methyl-4-(2,6,6-trimethylcyclohex-1-enyl) but-2-en-1-ol (3.67%), and 2-(2,4-hexadiynylidene)-1,6-dioxaspiro[4,4]non-3-ene (3.57%). Ninety-eight (93.83%) volatile flavor compounds were identified in the essential oil from the Aster yomena Makino. The major compounds were and 3-eicosyne (13.61%), 9,10,12-octadecatrienoic acid (7.8%), ${\alpha}$-caryophyllene alcohol (6.83%), 9-octadecynoic acid (6.03%), and ${\alpha}$-caryophyllene (5.74%). Although the two plants are apparently very similar, the chemical composition of the essential oils was significantly different in quality and quantity. In the case of C. zawadskii var. latilobum Kitamura, the sesquiterpene, valencene was found to be 10.82%, but it was not identified in A. yomena Makino. ${\delta}$-Cadinol appeared higher in C. zawadskii var. latilobum Kitamura than in A. yomena Makino. A clear characteristic of A. yomena Makino essential oil is that it has a high content of caryophyllene derivatives. The ${\alpha}$-caryophyllene alcohol contained in A. yomena Makino was relatively high at 6.83%, although the compound was not identified in C. zawadskii var. latilobum Kitamura. Also ${\alpha}$-caryophyllene was shown to be higher in A. yomena Makino than in C. zawadskii var. latilobum Kitamura.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.390-399
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• 2012

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, $pYES2M{\alpha}$-egIV, and $pYES2M{\alpha}$-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants $IpYES2M{\alpha}$-xegIV was higher than that of transformant IpYES2-xegIV or $IpYES2M{\alpha}$-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that $MF{\alpha}$ signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of $35^{\circ}C$ to $65^{\circ}C$. The optimal reaction condition for EGIV enzyme activity was at the temperature of $55^{\circ}C$, pH of 5.0, 0.75 mM $Ba^{2+}$, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant $IpYES2M{\alpha}$-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.

• 공업화학
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• 제5권5호
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• pp.779-785
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• 1994

양이온 색소인 thionin(Th)과 음이온 계면활성제인 sodium dodecyl sulfate(SDS) 사이의 상호작용을 흡수 spectra로 고찰하였다. 임계미셀농도(critical micelle concentration, CMC)보다 훨씬 낮은 premicellar 영역(S/D=10, 80 및 160)에서 온도가 증가함에 따라 몰 흡광계수비가 증가하거나 감소함을 볼 수 있으며, S/D=160에서 Th-SDS계의 다분자 회합체가 생성하는 가장 안정한 온도 범위는 $60^{\circ}C$ 이하임을 알 수 있었다. 한편 Th-SDS계에 첨가한 무기염 및 유기용매의 농도가 증가함에 따라 ${\gamma}$-band와 J-band는 감소하나 ${\alpha}$-band는 오히려 증가하는 경향이 나타났다. 이들의 ${\alpha}$-band의 증가 순서는 무기염의 경우 $Cl^-$>$ClO{_4}{^-}$>$SO{_4}{^{2-}}$>$NO{_3}{^-}$로 나타나며, 유기용매인 경우는 2-propanol>ethanol>methanol>ethylene glycol로 나타남을 알 수 있다.

• 한국미생물·생명공학회지
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• 제3권2호
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• pp.73-82
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• 1975

In the course of studies on the production of thermostable amylases by thermophilic actinomycetes isolated from soils the investigation was carried out on the production of $\alpha$-amylase by G-1011 strain which had presented the most remarkable $\alpha$-amylase formation ability among 128 amylolytic isolates. The results were as follows ： 1. Characteristics of G-1011 strain were compared with those descriptions of thermophilic actinomycetes given in Bergey's Manual. The strain was identical to these species of actinomycetes. The details of physiological properties of the strain luould he published in near future. 2. The optimum temperature for incubation of the cell growth of G-1011 strain and $\alpha$-amylase production by the strain was revealed to 5$0^{\circ}C$. 3. The effective medium for $\alpha$-amylase formation by the strain was consisted of 3.0%, soluble starch, 1.0%, peptone, 0.5%, yeast extract, 0.5%, NaCl, 0.1%, MgSO$_4$ㆍ7$H_2O$ 0.02%, $K_2$MPO$_4$and 0.002% FeSO$_4$ㆍ7$H_2O$. The pH of the medium was ajusted to 7.0 with phosphate buffer solution. 4. The maximum production of $\alpha$-amylase (3420 D. U/ml) by G-1011 strain resulted when it was grown for 16 hours with the culture of reciprocal shaking.

• 생약학회지
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• 제42권3호
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• pp.253-259
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• 2011

Macrophages play a vital role in the innate immune system involving defensive cytokines such as TNF (tumor necrosis factor)-${\alpha}$ and nitric oxide (NO). Therefore, we try to elucidate the anti-inflammatory activity of Chaga mushroom (Inonotus Obliquus, IO) in murine macrophages. Raw 264.7 cells and peritoneal macrophages of mice were cultured with or without LPS/LPS + IFN-${\gamma}$ in the presence of IO aqueous extracts (IOE 0.2, 2, 20, 100 ${\mu}g$/mL) for 24 hr and 48 hr, respectively. Exposure of IOE caused the decrease of NO production and increase of TNF-${\alpha}$ production in dose-dependent manner in activated peritoneal macrophage in vitro. To further investigate anti-inflammatory effects of IO ex vivo, we orally administrated capsaicin (PC, 3 mg/kg/day) and IOE (100, 200, 400 mg/kg/day) for 4 consecutive days to C57BL/6 mice (7~9 weeks old, female), then observed the NO secretion and cytokine (TNF-${\alpha}$) production of LPS/LPS + INF-${\gamma}$-stimulated peritoneal macrophages. IOE inhibits NO secretion in dose-dependent manner both ex vivo and in vitro and increases the production of TNF-${\alpha}$ in vitro. In addition, we found that IOE possessed suppressive effects of LPS-stimulated TNF-${\alpha}$, IL-$1{\beta}$, COX-2, as well as iNOS expressions in Raw 264.7 cells. These findings indicate that IOE suppress not only the LPS-induced NO overproduction of murine peritoneal macrophages, but also iNOS, COX-2, TNF-${\alpha}$, and IL-$1{\beta}$ overexpression of LPS-induced Raw 264.7 cells. Consequently, our results suggest that IO may have the anti-inflammatory effects via suppression of the inflammatory cytokines and mediators, and be useful for the treatment of inflammatory diseases.

• 대한화학회지
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• 제44권6호
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• pp.513-517
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• 2000

TTF (tetrathiafulvalene) 유도체의 전구체로서 5,6-dimethyl-1,3-dithiolo[4,5-b][1,4]dithiin-2-thione (화합물 3)을 로손반응에 의하여 합성하였다. 치환기의 종류에 따라 1,4-dithiin이나 티오펜이 포함된 두 가지의 서로 다른 생성물이 얻어졌고, 이들은 $^{13}C$ NMR과 고 분해능 전자충돌(HREI) 질량분석 분광학으로 특성화하였다. 화합물 3의 합성은 X-선 구조분석으로도 확인되었다. 결정학적 자료: 삼사정계, 공간군 P1, a=4.145(2)${\AA}$, b=10.600(2)${\AA}$, c=12.279(2)${\AA}$, ${\alpha}=71.440(10)^{\circ},\;{\beta}=84.30(2)^{\circ},\;{\gamma}=87.31(2)^{\circ}$, Z=2, $R(wR_2$)=0.0559(0.1416). 두 가지 생성물의 생성 기구를 비교하여 설명하였다.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.74-74
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• 2002

Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

• Tuberculosis and Respiratory Diseases
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• 제48권2호
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• pp.166-179
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• 2000

연구배경: 현재 사용되고 있는 많은 항암제는 암세포의 apoptosis를 유도함으로서 암세포를 사멸시키는 것이 주된 작용 기전이다. 항암제에 의한 치료 실패의 주된 원인은 암세포의 apoptosis에 대한 내성 획득에 의한 것으로 이해되고 있다. 최근의 보고에 의하면 많은 암세포주에서 apoptosis에 대한 내성 획득에 NF-${\kappa}B$의 활성화가 중요한 역할을 한다고 알려져 있지만 일부 세포에서는 상반된 결과를 보이고 있어 apoptosis에 대한 내성 획득과 NF-${\kappa}B$의 활성화와의 관련성은 세포에 따라 차이를 보이고 있다. 본 연구에서는 비소세포폐암 세포주가 TNF-$\alpha$ 유발 apoptosis에 저항을 나타내는 기전에서 NF-${\kappa}B$의 역할을 규명하고자 하였다. 방 법: 비소세포폐암세포주인 NCI-H157 세포에 TNF-$\alpha$ cycloheximide(CHX), TNF-$\alpha$와 CHX를 각각 투여하고 24시간 후 MTT assay로 세포생존율을 평가하였고 apoptosis의 발생 유무는 PARP에 대한 Westrn 분석으로 평가하였다. Ad5LacZ와 $Ad5I{\kappa}B{\alpha}SR$를 20시간 동안 감염시킨 각각의 세포를 1, 5, 10, 20, 50 ng/ml의 TNF-$\alpha$로 24, 48시간 자극 후 MTT assay를 시행하였고, 같은 농도의 TNF-$\alpha$로 24, 48시간 자극 후 PARP에 대한 Western 분석을 시행하였다. $I{\kappa}B{\alpha}$의 분해를 억제하는 proteasome inhibitor 인 MG132를 전처치하고 TNF-$\alpha$로 24, 48 시간 자극 후 MTT assay와 PARP에 대한 Westrn 분석을 시행하였다. $Ad5I{\kappa}B{\alpha}SR$를 감염시킨 세포를 TNF-$\alpha$로 자극한 후 NF-${\kappa}B$의 활성화를 EMSA로 평가하였다. 결 과: 1. TNF-$\alpha$ 단독 투여로는 세포 생존율의 감소와 apoptosis가 관찰되지 않았고 TNF-$\alpha$와 CHX를 같이 투여하였을 때는 세포 생존율의 감소와 함께 apoptosis가 유도되었다. 2. $Ad5I{\kappa}B{\alpha}SR$의 감염으로 $I{\kappa}B{\alpha}$가 과발현된 세포와 MG132를 전처치한 세포에서 TNF-$\alpha$ 자극에 의한 NF-${\kappa}B$의 활성화가 억제되었다. 3. $Ad5I{\kappa}B{\alpha}$ SR를 감염시킨 세포와 MG132를 전처치하여 NF-${\kappa}B$의 활성을 억제한 세포에서 TNF-$\alpha$ 자극 후 세포 생존율이 감소하고 apoptosis가 유도되었다. 결 론: 비소세포폐암 세포주의 TNF-$\alpha$ 유발 apoptosis에 대한 내성은 NF-${\kappa}B$의 활성화에 의해 생성되는 새로운 단백의 발현에 의한 것으로 생각되며, NF-${\kappa}B$ 활성화의 억제로 이를 극복할 수 있을 것으로 사료된다.

• Molecules and Cells
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• 제39권3호
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• pp.266-279
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• 2016

The mechanism by which 12-O-tetradecanoylphorbol-13-acetate (TPA) bypasses cellular senescence was investigated using human diploid fibroblast (HDF) cell replicative senescence as a model. Upon TPA treatment, protein kinase C (PKC) ${\alpha}$ and $PKC{\beta}1$ exerted differential effects on the nuclear translocation of cytoplasmic pErk1/2, a protein which maintains senescence. $PKC{\alpha}$ accompanied pErk1/2 to the nucleus after freeing it from $PEA-15pS^{104}$ via $PKC{\beta}1$ and then was rapidly ubiquitinated and degraded within the nucleus. Mitogen-activated protein kinase docking motif and kinase activity of $PKC{\alpha}$ were both required for pErk1/2 transport to the nucleus. Repetitive exposure of mouse skin to TPA downregulated $PKC{\alpha}$ expression and increased epidermal and hair follicle cell proliferation. Thus, $PKC{\alpha}$ downregulation is accompanied by in vivo cell proliferation, as evidenced in 7, 12-dimethylbenz(a)anthracene (DMBA)-TPA-mediated carcinogenesis. The ability of TPA to reverse senescence was further demonstrated in old HDF cells using RNA-sequencing analyses in which TPA-induced nuclear $PKC{\alpha}$ degradation freed nuclear pErk1/2 to induce cell proliferation and facilitated the recovery of mitochondrial energy metabolism. Our data indicate that TPA-induced senescence reversal and carcinogenesis promotion share the same molecular pathway. Loss of $PKC{\alpha}$ expression following TPA treatment reduces pErk1/2-activated SP1 biding to the $p21^{WAF1}$ gene promoter, thus preventing senescence onset and overcoming G1/S cell cycle arrest in senescent cells.