• Asian-Australasian Journal of Animal Sciences
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• v.28 no.4
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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).

• The Korean Journal of Food And Nutrition
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• v.31 no.3
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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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• v.22 no.3
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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.

• Applied Chemistry for Engineering
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• v.5 no.5
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• pp.779-785
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• 1994

The interaction between the cationic dye, thionin(Th) and the anionic surfactant, sodium dodecyl sulfate (SDS) has been investigated by absorption spectra. As the temperature of surfactant solution was increased in premicellar range(S/D=10, 80, and 160) which was much lower than the critical micelle concentration(CMC), the increment or decrement of the molar extinction coeffecient ratio appeared. It was found that the most stable temperature range of the oligomer aggregate in Th-SDS system at S/D=160 was below $60^{\circ}C$. With increasing the concentration of inorganic salt and organic solvents in Th-SDS system, ${\alpha}$-band was increased, but ${\gamma}$=band or J-band was decreased. The orders of ${\alpha}$-band increasing power were $Cl^-$>$ClO{_4}{^-}$>$SO{_4}{^{2-}}$>$NO{_3}{^-}$ and 2-propanol>ethanol>methanol>ethylene glycol.

• Microbiology and Biotechnology Letters
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• v.3 no.2
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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.

• Korean Journal of Pharmacognosy
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• v.42 no.3
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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.

• Journal of the Korean Chemical Society
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• v.44 no.6
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• pp.513-517
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• 2000

As a precursor of tetrathiafulvalene (TTF) derivative, 5,6-dimethyl-1,3-dithiolo[4,5-b][1,4]dithiin-2-thione (compound 3) was synthesized by the unusual Lawesson's reaction. Depending upon the substituents such as dimethyl and diphenyl groups, two different products containing 1,4-dithiin and thiophene moieties, respectively, were obtained and characterized by $^{13}C$ NMR and high-resolution electron impact (HREI) mass spectroscopy. The formation of 3 was further characterized by X-ray structure analysis. Crystallographic data for 3: triclinic, space group 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 and R(wR$_2$)=0.0559(0.1416). The formation mechanism of two products was suggested and compared each other.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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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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• v.48 no.2
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• pp.166-179
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• 2000

Background: The main reason for the failure of anti-cancer chemotherapy is the build up of resistance by cancer cells to apoptosis. The activation of NF-${\kappa}B$ in many cancer cell lines is reported to be underlying mechanism behind the build up of resistance of cancer cells to apoptosis. However, this relationship varied depending on the cells used in the experiments. In this study, the role of NF-${\kappa}B$ activation in the TNF-$\alpha$-induced apoptosis in lung cancer cell line was evaluated. Methods: NCI-H157 cells were used in all experiments. Cells were exposed to a high dose of TNF-$\alpha$(20 ng/ml) for 24 or 48 hours with or without blocking NF-${\kappa}B$ activation. TNF-$\alpha$-induced activation of NF-${\kappa}B$ was inhibited either by overexpression of $I{\kappa}B{\alpha}$-super repressor($I{\kappa}B{\alpha}$-SR) or by pre-treatment with proteasome inhibitor. Cell viability and apoptosis were evaluated with MTT assay and Western blot analysis for PARP fragment, respectively. Results: Cell viability of NCI-H157 cells was not affected by TNF-$\alpha$ treatment alone; however, combined treatment with TNF-$\alpha$ and cycloheximide reduced cell viability significantly, indicating that resistance to TNF-$\alpha$ is mediated by the new proteins synthesized after TNF-$\alpha$ stimulation. To evaluate the role of NF-${\kappa}B$ in the transcription of anti-apoptotic proteins. delete NF-${\kappa}B$ activation was inhibited before TNF-$\alpha$ stimulation. as described above. $AD5I{\kappa}B{\alpha}$-SR-transduction inhibited TNF-$\alpha$-induced nuclear translocation of p65. TNF-$\alpha$-induced cell death and apoptosis increased after inhibition of TNF-$\alpha$-induced activation of NF-${\kappa}$ by methods. Conclusion: These results suggest that TNF-$\alpha$-induced activation of NF-${\kappa}B$ may be closely related to the acquisition of the resistance to TNF-$\alpha$-induced apoptosis in lung cancer cells. Therefore. blocking of NF-${\kappa}B$ pathway can be a useful therapeutic modality in the treatment of lung cancer.

• Molecules and Cells
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• v.39 no.3
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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.