• Title/Summary/Keyword: Reaction factor

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Expression of vascular endothelial growth factor in oral squamous cell carcinoma

  • Kim, Seok-Kon;Park, Seung-Goo;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.41 no.1
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    • pp.11-18
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    • 2015
  • Objectives: The goal of this study was to determine the correlation of clinicopathological factors and the up-regulation of vascular endothelial growth factor (VEGF) expression in oral squamous cell carcinoma. Materials and Methods: Immunohistochemical staining of VEGF and quantitative real-time polymerase chain reaction (RT-PCR) of VEGF mRNA were performed in 20 specimens from 20 patients with oral squamous cell carcinoma and another 20 specimens from 20 patients with carcinoma in situ as a controlled group. Results: The results were as follows: 1) In immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, high-level staining of VEGF was observed. Significant correlation was observed between immunohistochemical VEGF expression and histologic differentiation, tumor size of specimens (Pearson correlation analysis, significance r>0.6, P<0.05). 2) In VEGF quantitative RT-PCR analysis, progressive cancer showed more VEGF expression than carcinoma in situ. Paired-samples analysis determined the difference of VEGF mRNA expression level between cancer tissue and carcinoma in situ tissue, between T1 and T2-4 (Student's t-test, P<0.05). Conclusion: These findings suggest that up-regulation of VEGF may play a role in the angiogenesis and progression of oral squamous cell carcinoma.

Biochemical Characterization of a Putative Calcium Influx Factor as a Diffusible Messenger in Jurkat Cells, Xenopis Oocytes, and Yeast

  • Kim, Hak-Yong
    • Animal cells and systems
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    • v.7 no.1
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    • pp.75-79
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    • 2003
  • Highly purified high performance thin layer chromatography (HPTLC) fractions containing a putative calcium influx factor (CIF) were prepared from the Jurkat cells and Xenopus oocytes in which $Ca^{2+}$ stores were depleted by thapsigargin treatment and from the yeast in which intracellular $Ca^{2+}$ stores were also depleted by genetic means. Microinjection of the fractions has been shown to elicit $Ca^{2+}$ dependent currents in Xenopus oocytes. The nature of the membrane currents evoked by the putative CIF appeared to be carried by chloride ions since the current was blocked by the selective chloride channel blocker 1 mM niflumic acid and its reversal potential was about -24 mV. Injection of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid (BAPTA) eradicated the current activities, suggesting the current responses are entirely $Ca^2$-dependent. Moreover, the currents were sensitive to the removal of extracellular calcium, indicating the dependence on calcium entry through the plasma membrane calcium entry channels. CIF activities were insensitive to protease, heat, and acid treatments and to Dische-reaction whereas the activities were sensitive to nucleotide pyrophosphatase and hydrazynolysis. The fraction might have a sugar because it was sensitive to Molisch test and Seliwaniff's resorcinol reaction. From the above results, CIF as a small and stable molecule seems to have pyrimidine, pyrophosphate, and a sugar moiety.oiety.

Molecular Characterization and Expression Analysis of Equine Vascular Endothelial Growth Factor Alpha (VEGFα) Gene in Horse (Equus caballus)

  • Song, Ki-Duk;Cho, Hyun-Woo;Lee, Hak-Kyo;Cho, Byung Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.5
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    • pp.743-748
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    • 2014
  • The objective of this study was to determine the molecular characteristics of the horse vascular endothelial growth factor alpha gene ($VEGF{\alpha}$) by constructing a phylogenetic tree, and to investigate gene expression profiles in tissues and blood leukocytes after exercise for development of suitable biomarkers. Using published amino acid sequences of other vertebrate species (human, chimpanzee, mouse, rat, cow, pig, chicken and dog), we constructed a phylogenetic tree which showed that equine $VEGF{\alpha}$ belonged to the same clade of the pig $VEGF{\alpha}$. Analysis for synonymous (Ks) and non-synonymous substitution ratios (Ka) revealed that the horse $VEGF{\alpha}$ underwent positive selection. RNA was extracted from blood samples before and after exercise and different tissue samples of three horses. Expression analyses using reverse transcription-polymerase chain reaction (RT-PCR) and quantitative-polymerase chain reaction (qPCR) showed ubiquitous expression of $VEGF{\alpha}$ mRNA in skeletal muscle, kidney, thyroid, lung, appendix, colon, spinal cord, and heart tissues. Analysis of differential expression of $VEGF{\alpha}$ gene in blood leukocytes after exercise indicated a unimodal pattern. These results will be useful in developing biomarkers that can predict the recovery capacity of racing horses.

Effects of confinement on physiological and psychological responses and expression of interleukin 6 and brain derived neurotrophic factor mRNA in primiparous and multiparous weaning sows

  • Zhang, Mingyue;Li, Xiang;Li, Jianhong;Sun, Hanqing;Zhang, Xiaohui;Bao, Jun
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.9
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    • pp.1350-1357
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    • 2017
  • Objective: The present study aimed to investigate whether the long-lasting, recurrent restricting of sows leads to the physiological and psychological reaction of discomfort. Methods: Sows (Large White) that had experienced restricting for about 0.5 or 3 years and agematched sows kept in a group housing system (loose sows) were compared. Pupillary light reflex parameters were measured at the weaning stage. Immediately after slaughter, blood samples were taken to measure serum cortisol levels, and the brain was dissected, gene expression in the hippo-campus, frontal cortex and hypothalamus was analyzed. Results: The serum cortisol levels were higher in the confined sows than in the loose sows. The full maturity, but not the young adolescent, confined sows had longer latency time in the onset of pupil constriction than their loose counterparts. Real-time polymerase chain reaction analyses revealed an increased expression of interleukin 6 mRNA in the hippocampus and decreased expression of brain derived neurotrophic factor mRNA in hippocampus and hypothalamus and to a lesser extent in the frontal cortex of the full maturity confined sows, compared with the full maturity loose sows. Conclusion: Taken together, these data indicated that recurrent restricting stress in full maturity sows leads to the physiological and psychological reaction of discomfort.

Effect of Water-impregnation on Steam Explosion of Pinus densiflora

  • Jung, Ji Young;Ha, Si Young;Yang, Jae-Kyung
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.2
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    • pp.189-199
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    • 2019
  • This study addresses the biorefinery feedstock from Pinus densiflora. This raw material is a major tree species in the Republic of Korea; it is renewable, has cost-effective, and is readily available. In this study, steam explosion of P. densiflora was performed in a reactor at $225^{\circ}C$ and with 1 to 13 min reaction times with or without previous water impregnation. The combined severity factor (Ro), which is an expression relating the reaction temperature and reaction time used in the steam explosion treatment, ranged from 3.68 to 4.79. The influence of both impregnation and steam explosion conditions were investigated by examining color variations, chemical composition, and mass balance on the pretreated solids. The results showed that steam-exploded P. densiflora that was not impregnated with water exhibited significantly darker color (chroma 28.8-41.4) than water-impregnated and steam-exploded P. densiflora (chroma 18.8-37.3). The increased ${\alpha}$-cellulose and lignin contents were detected as the severity factor increased. Furthermore, the ${\alpha}$-cellulose and lignin contents in the non-impregnated/steam-exploded P. densiflora were higher than those in the water-impregnated/steam-exploded P. densiflora. However, the decreased holocellulose content was detected as the severity factor increased. In mass balance, the holocellulose yield from water-impregnated/steam-exploded P. densiflora was higher than that from the non-impregnated P. densiflora.

Association of single-nucleotide polymorphisms in dual specificity phosphatase 8 and insulin-like growth factor 2 genes with inosine-5'-monophosphate, inosine, and hypoxanthine contents in chickens

  • Jean Pierre Munyaneza;Minjun Kim;Eunjin Cho;Aera Jang;Hyo Jun Choo;Jun Heon Lee
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1357-1366
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    • 2023
  • Objective: This study aimed to identify the single-nucleotide polymorphisms (SNPs) in the dual-specificity phosphatase 8 (DUSP8) and insulin-like growth factor 2 (IGF2) genes and to explore their effects on inosine-5'-monophosphate (IMP), inosine, and hypoxanthine contents in Korean native chicken -red-brown line (KNC-R Line). Methods: A total sample of 284 (males, n = 127; females n = 157) and 230 (males, n = 106; females, n = 124) aged of 10 weeks old KNC-R line was used for genotyping of DUSP8 and IGF2 genes, respectively. One SNP (rs313443014 C>T) in DUSP8 gene and two SNPs (rs315806609A/G and rs313810945T/C) in IGF2 gene were used for genotyping by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and KASP methods, respectively. The Two-way analysis of variance of the R program was used to associate DUSP8 and IGF2 genotypes with nucleotide contents in KNC-R chickens. Results: The DUSP8 (rs313443014 C>T) was polymorphic in KNC-R line and showed three genotypes: CC, CT, and TT. The IGF2 gene (rs315806609A/G and rs313810945T/C) was also polymorphic and had three genotypes per SNP, including GG, AG, and AA for the SNP rs315806609A/G and genotypes: CC, CT, and TT for the SNP rs313810945T/C. Association resulted into a strong significant association (p<0.01) with IMP, inosine, and hypoxanthine. Moreover, the significant effect of sex (p<0.05) on nucleotide content was also observed. Conclusion: The SNPs in the DUSP8 and IGF2 genes might be used as genetic markers in the selection and production of chickens with highly flavored meat.

Monitoring of Dynamic Changes in Maillard Reaction Substrates by Response Surface Methodology (반응표면분석에 의한 Maillard 반응기질의 동적변화 모니터링)

  • Lee, Gee-Dong;Kim, Jeong-Sook;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
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    • v.28 no.2
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    • pp.212-219
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    • 1996
  • Four-dimensional response surface methodology was used for monitoring dynamic changes in substrates during Maillard reaction. The coefficients of determination ($R^2$) of response surface regression equations for the changes in amino acids during Maillard reaction were 0.9478 for total amino acids and above 0.90 for each amino acid. $R^2$ of regression equations for the changes in sugars during Maillard reaction were 0.9250 for glucose and 0.6490 for fructose. The contents of total amino acids gradually decreased with increasing reaction temperature and pH of the solvent. Browning color intensity increased with rising reaction temperature, showing maximum color intensity at around $145^{\circ}C$. Each amino acid showed a decreasing tendency in its contents, which was similarly found in total amino acids. Four-dimensional response surface methodology indicated that the increased temperature during Maillard reaction was the most influential factor in decreasing substrates, such as aspartic acid, threonine and glucose. While the reaction time and pH of solvent little affected the changes in the above-mentioned substrates during Maillard reaction.

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Cloning of Bovine Macrophage Colony-stimulating Factor

  • Kim, Tae-Yung;Kim, Cheol-Ho;Lee, Sang-Gil;Kang, Chung-Boo
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.892-897
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    • 2005
  • Macrophage colony-stimulating factor (M-CSF) is a growth factor required for growth and differentiation of mononuclear phagocyte lineage. Total and 16 poly (A) mRNA of bovine M-CSF were isolated from healthy bovine peripheral mononuclear cells stimulated by phobol 12-myristste 13-acetate (TPA). The more compatible cultured mononuclear cells were 5${\times}$10/ml for RNA isolation. TPA-activated mononuclear cells increased the level of M-CSF-mRNA more than concanavalin A (Con A) and lipopolysaccharide (LPS). The optimal analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) for14 Macrophage colonystimulating factor (M-CSF) as a growth factor required for bovine M-CSF was denaturation at 94$^{\circ}C$ for 1 minute, annealing at 57$^{\circ}C$ for 1 minute, extension at 72$^{\circ}C$ for 1 minute for 30 cycles. The size of cDNA of bovine M-CSF by RT-PCR was 774 base pairs. A 774 base pairs cDNA encoding bovine M-CSF was synthesized by reverse transcriptase polymerase chain reaction (RT-PCR). Ligated cDNA was transformed to competent cells and then plasmid isolation and digestion was performed. Molecular cloning and sequencing were performed for cDNA of bovine M-CSF. The size of cloned cDNA of bovine M-CSF was 774base pairs. The homology of base sequence and amino acid sequence was 88% and 86% compared with known human M-CSF, respectively. From a high degree of sequence similarity, the obtained cDNA of bovine M-CSF is thought be a specific gene of bovine M-CSF.

Optimization of One-step Extraction/Methylation Method for Analysis of Fatty Acid Composition in Brown Rice

  • Cho, Kyoung-Shim;Kim, Hyun-Ju;Moon, Sang-Mi;Kang, Jung-Hoon;Lee, Young-Sang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.1
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    • pp.89-94
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    • 2006
  • Traditionally fatty acid composition used to be analysed by a GC and the sample preparation process includes lipid extraction from sample and subsequent methyl esters preparation, which are time-consuming and cumbersome. As an alternative, simultaneous extraction/methylation methods are being developed for rapid and simplified sample preparation. To optimize one-step extraction/methylation method for analysis of fatty acid composition in brown rice, various reaction factors such as sample to reaction solution ratio, reaction time and temperature, shaking intensity were changed and resultant fatty acid composition data were evaluated in comparison with previous reports. The ratio of sample weight to reaction solution volume was the most critical factor in that higher sample to reaction solution ratio caused overestimation of palmitic acid and linoleic acid composition, resulting in underestimation of oleic acid. Lower reaction temperature also induced overestimation of linoleic acid and underestimation of oleic acid. Reaction duration and the intensity of shaking prior to and during the reaction, however, caused no significant changes in analysis results. In conclusion, the optimum condition was mixing 5 grains (about 0.2 g) of brown rice with $680{\mu}L$ of extraction/methylation mixture and $400{\mu}L$ of heptane, followed by reaction at $80^{\circ}C$ for 2 hours.

Effect of Extracellular $Ca^{2+}$ and $Ca^{2+}$-ATPase on the Acrosome Reaction of Spermatozoa (세포외 $Ca^{2+}$$Ca^{2+}$-ATPase가 정자의 첨체반응에 미치는 영향)

  • Yung-Keun Oh;Jae-Ho Chang;In-Ho Choi;Noh-Pal Jung;Hyung-Cheul Shin;Byoung-Ju Kwak
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.27-33
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    • 1998
  • This study has been designed in order to examine a physiological role of $Ca^{2+}$ which has been known as an essential factor for capacitation, to confirm whether the enzyme activity of $Ca^{2+}$-ATPase on capacitation is important or not, and to clarify relationship between various levels of the $Ca^{2+}$ concentration and $Ca^{2+}$-ATPase which has been known to be an important factor of the plasma membranes. In the present study applying quercetin, a $Ca^{2+}$-ATPase inhibitor, the enzymatic effect of $Ca^{2+}$-ATPase on capacitation was found to be remarkable: a significant increase of the transition from the original type (type A) to the type B and the type AR of the spermatozoa. This finding suggests that $Ca^{2+}$-ATPase plays an important role in the efflux and the influx of the $Ca^{2+}$ which has been known to be an essential factor the capacitation and acrosome reaction, and that the inhibitory action of the $Ca^{2+}$-ATPase might be a prerequsite step toward the acrosome reaction. The conclusion reached can be deduced as follows: increment of the intracelluar $Ca^{2+}$ concentration occurred by controlling the slope of $Ca^{2+}$ concentration through $Ca^{2+}$-ATPase activities in both the intra- and extracelluar fluid may be an important procedure for capacitation and acrosome reaction, and ultimately for fertilization of the spermatozoa and the ova.

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