• Title/Summary/Keyword: Protein p16

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Proteomics Approach on Puroindoline Gene of Pre-harvest Sprouting Wheat

  • Kamal, Abu Hena Mostafa;Park, Cheol-Soo;Heo, Hwa-Young;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Woo, Sun-Hee
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.205-212
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    • 2009
  • Wheat (Triticum aestivum L.) grain texture is an important determinant of milling properties and end product use. Two linked genes, puroindoline a (PINA) and puroindoline b (PINB), control most of the genetic variation in wheat grain texture. Wheat seed proteins were examined to identify PINA and PINB gene using two pre-harvest sprouting wheat cultivars; Jinpum (resistant) and Keumgang (susceptible).Wheat seed proteins were separated by two-dimensional electrophoresis with IEF gels over pH ranges: pH 3-10. A total of 73 spots were digested with trypsin resulting peptide fragmentation were analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS). Mass spectra were automatically processed and searched through NCBInr, SWISS-PORT and MSDB database with mono isotopic masses and complete gene sequence were found by UniProt database. Puroindoline a and puroindoline b that is responsible for grain texture related with baking performance and roughness. Two spots were found Pin b (16.7 kDa) and Pin a (16.3 kDa) in Jinpum compare to seven spots were identified Pin a (16.1 kDa, 16.3 kDa) and Pin b (16.7 kDa, 9.5 kDa and 14.4 kDa) in Keumgang. Some selected spots were identified puroindoline like grain softness protein (16.9 kDa, 17 kDa and 18.1 kDa) in Keumgang. Moreover, to gain a better inferring the identification of puroindoline related proteins using proteomics, we accomplished a complete gene sequence of PINA and PINB gene in pre-harvesting sprouting wheat seeds between resistant (Jinpum) and susceptible (Keumgang).

Effects of an Extract from the Roots of Platycodon Grandiflorum on the Levels of p53 and pRB in NCI-H460 Human Lung Carcinoma Cells (길경 수용액 추출물에 의한 NCI-H460 인체 폐암세포의 p53 및 pRB의 발현에 미치는 영향)

  • Park, Bong-Kyu;Gam, Chul-Woo;Heo, Tae-Yool;Park, Dong-Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1530-1537
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    • 2006
  • Platycodi Radix, the root of Platycodon grandiflorum A. DC (Campanulaceae), commonly known as Doraji in Korea (Chinese name, 'Jiegeng', and Japanese name, 'Kikyo') has been used as an expectorant in traditional Oriental medicine. Extracts from the roots of P. grandiflorum have been reported to have wide ranging health benefits. In Korea, Platycodi Radix is also used as a food and employed as a folk remedy for adult diseases, such as bronchitis, asthma and pulmonary tuberculosis, hyperlipidemia, diabetes, and inflammatory diseases, and as a sedative. Several studies on its chemical and immunopharmacological effects including immunostimulation and antitumor activity have been performed. However, the relevant molecular mechanisms are poorly understood. Platycodi Radix, the root of Platycodon grandiflorum A. DC (Campanulaceae), commonly known as Doraji in Korea (Chinese name, 'Jiegeng', and Japanese name, 'Kikyo') has been used as an expectorant in traditional Oriental medicine. Extracts from the roots of P. grandiflorum have been reported to have wide ranging health bensfits. In Korea, Platycodi Radix is also used as a food and employed as a folk remedy for adult diseases, such as bronchitis, asthma and pulmonary tuberculosis, hyperlipidemia, diabetes, and inflammatory diseases, and as a sedative. Several studies on its chemical and immunopharmacological effects including immunostimulation and antitumor activity have been performed. However, the relevant molecular mechanisms are poorly understood. In the present study, we investigated the effects of an aqueous extract from the roots of P. grandiflorum AEPG) on the cell growth of human lung adenocarcinoma NCI-H460 cells in order to understand its anti-proliferative mechanism. AEPG treatment down-regulated the cyclin D1 expression in both transcriptional and translational levels without alteration of cyclin E. In AEPG-treated cells, the levels of cyclin-dependent kinase (C아) 6 mRNA and protein were significantly inhibited, but the levels of Cdk2 and Cdk4 were slightly inhibited by treatment of AEPG. AEPG treatment induced a marked accumulation of Cdk inhibitors, p16 and p27. However, AEPG treatment did not affect not only retinoblastoma protein (pRB) but also tumor suppressor p53 protein expression. The present results indicated that AEPG-induced inhibition of lung cancer cell proliferation is associated with the blockage of G1 phase progression through induction of Cdk inhibitors such as p16 and p27, and inhibition of cyclin D1 and Cdk6. AEPG exposure, as offered by this study, provides cluse for the mechanism of AEPG action. Taken together, these findings suggest that P. grandiflorum has strong potential for development as an agent for prevention and treatiment against human lung cancer.

Development of SSLP Marker Targeted to P34 Null Gene in Soybean (콩 P34 단백질 결핍 유전자를 이용한 SSLP 마커 개발)

  • Yang, Kiwoung;Ko, Jong-Min;Lee, Young-Hoon;Jeon, Myeong Gi;Jung, Chan-Sik;Baek, In-Youl;Kim, Hyun-Tae;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.502-506
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    • 2010
  • Soybean seed possesses about 15 allergenic proteins recognized by IgEs from soy-sensitive human. The allergenic impact of soybean proteins limit its extensive usage in a broad range of processed foods. Soybean protein P34 or Gly m Bd 30k of the cysteine protease family is one of the major allergen of the soybean seed. P34-null soybean, PI567476, was identified among soybean (Glycine max & Glycine soja Sieb. and Zucc) of approximately 16,226 accessions from USDA soybean germplasm screened. Also, for P34 gene (Williams 82; whole genome sequence cultivar) and P34 null gene (PI567476) comparative analysis of sequences listed in the NCBI database showed the presence of a SSLP (Simple Sequence Length Polymorphism) of 4 base pair. So, a SSLP marker was designed to reveal the polymorphism of the locus. In this study, a population of 339 $F_2$ recombinant inbred lines generated by cross between Taekwang (Glycine max) and PI567476 was used to select $F_{2:3}$ plant of a P34 null gene. The result separation rate Taekwang type, heterozygous type and PI567476 type were shown in 85: 187: 67 since single gene is concerned in as the separation rate of 1:2:1 in $X^2{_{0.05}}=5.99$, df=2. In future, selected plant will identify protein level, whether P34 null protein is equal to P34 null gene.

Purification and Characterization of $L-galactono-{\gamma}-lactone$ Oxidase in Pichia sp. Isolated from Kimchi (김치유래 Pichia속 효모가 생산하는 $L-galactono-{\gamma}-lactone\;oxidase$의 분리 정제 및 특성)

  • Oh, Ji-Young;Han, Young-Sook
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1135-1142
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    • 2003
  • The purification and characteristics of the biosynthesis enzyme of vitamin C from microorganisms related with kimchi fermentation were investigated to define vitamin C biosynthetic pathways in yeast. A yeast strain (Pichia onychis 16-4) which synthesizes vitamin C with galacturonic acid as substrate at high rate was isolated from kimchi. The enzyme $L-galactono-{\gamma}-lactone$ oxidase isolated from the yeast was purified and characterized. The specific activity of the crude enzyme was 7.26 unit/mg protein, which increased to 4,698 unit/mg protein with a chromatography of Sephacryl S-200HR; indicating a 647.1-fold level of purification. The molecular weights of the dissociated enzymes were estimated to be 31,000, 39,000, and 50,000 KD. Among the substrates tested, $L-galactono-{\gamma}-lactone$ was the most effective. The enzyme showed optimum activity ah pH 7.8 and 35c. The purified enzyme uses $O_2$ as the electron acceptor for oxidation of $L-galactono-{\gamma}-lactone$.

The Effects of Caponization Age on Muscle Characteristics in Male Chicken

  • Chen, Kuo-Lung;Chen, Tsai-Tzu;Lin, Kou-Joong;Chiou, Peter Wen-Shyg
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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    • pp.1684-1688
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    • 2007
  • This study examined the caponization effects on the muscle characteristics (quality and quantity) of caponized male chickens fed before or after sexual maturity. Healthy and uniform Single Comb White Leghorn chickens were caponized at 3-week-old. Feeding was conducted until 16-week-old in trial 1 or from 12-week-old to 26-week-old in trial 2. Ten sham operated male chickens (Sham) were also assigned to each trial as the control group. Chickens used in both trials were housed in individual cages with each chicken representing one replicate. The results showed that early caponization (3-week-old) significantly increased (p<0.05) body weight and pectoral major muscle weight and percentage at 16-week-old compared to the Sham in trial 1. Caponization significantly increased (p<0.05) the protein content of the pectoral major muscle, but decreased (p<0.05) the ash content. Late caponization (12-week-old) significantly decreased (p<0.05) the ash content, myofibrillar ATPase activity and emulsification capacity of the pectoral major muscle in mature capons (26-week-old) compared to the Sham in trial 2. Early caponization (3-week-old) only increased the weight and protein content of the pectoral major muscle with decreased ash content in 16-week-old capons. Late caponization (12-week-old) showed no affects on pectoral major muscle quantity, while it decreased the ATPase activity and enhanced the emulsification capacity in mature (26-week-old) capons. Hence, the muscle quality improvement was shown as capons were fed to sexual maturity.

Insulin-like Growth Factor-1, IGF-binding Protein-3, C-peptide and Colorectal Cancer: a Case-control Study

  • Joshi, Pankaj;Joshi, Rakhi Kumari;Kim, Woo Jin;Lee, Sang-Ah
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3735-3740
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    • 2015
  • Context: Insulin-like growth factor peptides play important roles in regulating cell growth, cell differentiation, and apoptosis, and have been demonstrated to promote the development of colorectal cancer (CRC). Objective: To examine the association of insulin-related biomarkers including insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein-3 (IGFBP-3) and C-peptide with CRC risk and assess their relevance in predictive models. Materials and Methods: The odds ratios of colorectal cancer for serum levels of IGF-1, IGFBP-3 and C-peptide were estimated using unconditional logistic regression models in 100 colorectal cancer cases and 100 control subjects. Areas under the receiving curve (AUC) and integrated discrimination improvement (IDI) statistics were used to assess the discriminatory potential of the models. Results: Serum levels of IGF-1 and IGFBP-3 were negatively associated with colorectal cancer risk (OR=0.07, 95%CI: 0.03-0.16, P for trend <.01, OR=0.06, 95%CI: 0.03-0.15, P for trend <.01 respectively) and serum C-peptide was positively associated with risk of colorectal cancer (OR=4.38, 95%CI: 2.13-9.06, P for trend <.01). Compared to the risk model, prediction for the risk of colorectal cancer had substantially improved when all selected biomarkers IGF-1, IGFBP-3 and inverse value of C-peptide were simultaneously included inthe reference model [P for AUC improvement was 0.02 and the combined IDI reached 0.166% (95 % CI; 0.114-0.219)]. Conclusions: The results provide evidence for an association of insulin-related biomarkers with colorectal cancer risk and point to consideration as candidate predictor markers.

Diagnosis of Protein Losing Enteropathy in Connective Tissue Diseases with $^{99m}Tc$-human Serum Albumin(HSA) (교원병에 동반된 단백소실장질환(Protein Losing Enteropathy)에서의 테크네슘표지 인혈청알부민 스캔)

  • Won, Kyoung-Sook;Oh, Yeong-Seok;Bang, Shin-Ho;Park, Won
    • The Korean Journal of Nuclear Medicine
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    • v.27 no.1
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    • pp.88-97
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    • 1993
  • Anterior abdominal scintigraphy after intravenous injection of $^{99m}Tc$-human serum albumin ($^{99m}Tc$-HSA 20 mCi) was done in 16 patients with connective tissue diseases and 15 healthy control patients. Patients with proteinuria or hepatopathy were excluded. 1) 7 (44%) patients among 16 connective tissue disease patients without the apparent evidence of external protein loss showed abnormal intestinal accumulation of albumin. 6 patients with positive albumin scintigraphy showed bypoalbuminemia. 2) There was no false positive scintigraphic finding in control group. 3) The serum albumin level in connective tissue disease patients ($3.1{\pm}0.6 g/dl$, n=16) was lower than control patients ($3.9{\pm}0.3 g/dl$, n=15) (p<0.0001). 4) The serum albumin level was lower in connective tissue disease patients with positive $^{99m}Tc$-HSA scan ($2.8{\pm}0.6 g/dl$, n=7) than the connective tissue disease patients with negative scan ($3.3{\pm}0.3 g/dl$, n=9) (p<0.05). 5) The hemoglobin level in connective tissue disease patients with positive nan ($10.6{\pm}2.91 g/dl$) was lower than that of the control group ($13.6{\pm}1.5 g/dl$) (p<0.05). Mypoalbuminemia is frequently involved in chronic connective tissue diseases. Protein losing enteropathy (PLE) is also responsible for the majority of the bypoalbuminemia in these patients. But it has been ignored because the conventional method for the diagnosis of PLE was difficult to perform. $^{99m}Tc$-HSA scan also must be validated by more extended study and comparison with the quantitative study such as stool ${\alpha}-1$ antitrypsin measurement. There must be a reevaluation of PLE in various diseases especially in connective tissue diseases with easy, fast, economical, and non-invasive method.

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Cytokine mRNA Expression in the Small Intestine of Weanling Pigs Fed Diets Supplemented with Specialized Protein or Peptide Sources

  • Zhao, J.;Harper, A.F.;Webb, K.E. Jr.;Kuehn, L.A.;Gilbert, E.;Xiao, X.;Wong, E.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1800-1806
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    • 2008
  • Cytokines play a central role in the mucosal immune response and are involved in regulation of nutrient absorption, metabolism and animal growth. This study investigated the effect of diet manipulation with specialized protein or peptide sources on expression of cytokine (IL-1, IL-6, IL-10, and TNF-${\alpha}$) mRNA abundance in different intestinal regions and at different ages post-weaning in piglets. A total of 48 (17 days of age, $6.16{\pm}0.34kg\;BW$) weanling pigs were fed either a corn-soy/whey protein basal diet, the basal diet supplemented with spray-dried plasma protein (SDPP), or the basal diet supplemented with $Peptiva^{(R)}$, a hydrolyzed marine plant protein. A fourth treatment group was fed the SDPP diet, but the feed intake level was limited (SDPP-LF). Pigs were killed at 3 and 10 d, and intestinal cytokine mRNA was measured by real-time PCR using the relative quantification method. The SDPP-LF group exhibited an increased TNF-${\alpha}$ mRNA abundance compared with the ad libitum SDPP group (p<0.05). The TNF-${\alpha}$ and IL-10 mRNA abundance increased from the proximal to distal part of the intestine, and the mRNA abundance was greater (p<0.01) in the distal intestine as compared with the proximal and middle intestine. The cytokines IL-1-${\beta}$, IL-10 and TNF-${\alpha}$ mRNA abundance also increased from d3 to d10 postweaning (p<0.01). In summary, restricted feeding increased the TNF-${\alpha}$ mRNA abundance in the small intestine, however neither SDPP nor peptide supplementation affected cytokine mRNA expression. Abundance of mRNA for most cytokines examined in this study increased with age post-weaning, suggesting that during 10 d after weaning the mucosal immune system is still under development.

Effect of Potassium Chloride on the Separation and Purification of Model Proteins using Counter Current Distribution (역류분배에 의한 모델단백질의 분리정제시 염화칼륨의 영향에 관한 연구)

  • Lee, Chang-Ho;Lee, Boo-Yong;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.22 no.2
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    • pp.177-182
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    • 1990
  • The changes in the partition coefficient of model proteins (lysozyme, myoglobin, conalbumin, bovine serum albumin) in an aqueous two·phase system formed by polyethylene glycol and dextran were examined in order to improve the capacity of counter current distribution (CCD) for the protein fractionation and concentration . The protein distribution pattern in CCD with 30 tubes varied with the pH (4.5, 5.5, 6.5, 9.0, 12.0) and KCl concentration (0mM, 50mM, 250mM, 500mM) of the system. From the mixture of model proteins, pure myoglobin was appeared at the upperphase of 14th tube having 50mM of KCl at pH 5.5 and the upper-phase of 13th tube having 250mM of KCl at pH 6.5. Similarly pure BSA was obtained at the 14th tube having KCl 250mM with pH 4.5, pure lysozyme at the 19th tube having 500mM of KCl at pH 4.5 and the upper-phase of 16th tube 50mM of KCl at pH 5.5.

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Digestion and Nitrogen Utilization by Sheep Fed Diets Supplemented with Processed Broiler Litter

  • Kwak, W.S.;Fontenot, J.P.;Herbein, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.11
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    • pp.1634-1641
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    • 2003
  • In vivo digestion and metabolism trials were conducted with 10 wethers equipped with ruminal, abomasal, and ileal cannulae to evaluate digestion of ensiled broiler litter (EBL), deepstacked broiler litter (DBL), and composted broiler litter (CBL). Wethers were fed a low protein (6.3% CP) basal diet alone or supplemented to 10.3% CP with EBL, DBL, CBL or soybean meal (SBM). All diets were formulated to be isoenergetic (56% TDN, DM basis). Apparent digestibilities of DM, OM, and ADF were not affected (p<0.05) by diet, but digestibility of CP was improved (p<0.05) by N supplementation. Apparent digestibility of CP was lower (p<0.05) for diets supplemented with CBL and DBL than for diets supplemented with SBM and EBL. Ruminal $NH_3$ concentration was 20 to 24 mg/dl at 2 h after feeding litter-supplemented diets compared with 13 mg/dl for SBM. Abomasal N, $NH_3$ N, and nonammonia N flows were increased (p<0.05) by N supplementation, whereas microbial N flow was not influenced (p<0.05) by diet. Compared with SBM and EBL, undegraded dietary CP flow to the abomasum tended to be greater (p<0.1) when wethers were fed DBL and CBLsupplemented diets. Retention of N (g/d) also was greater (p<0.05) due to greater (p<0.05) N intake and lower (p<0.05) urinary N excretion when wethers were fed diets supplemented with litter (especially EBL) vs. SBM. Overall, characteristics of ruminal fermentation and digestion indicated that broiler litter N was utilized efficiently by wethers, but ensiling may be preferable to deepstacking or composting.