An experiment was carried out to evaluate the effects of feeding graded levels of chromium in the form of chromium picolinate on growth performance, blood components, carcass grade, in vitro lipogenesis and lipolysis, and in vitro protein degradation and synthesis in growing-finishing pigs. There were no significant differences for daily weight gain, feed intake and feed conversion among treatments during growing phase, while in the finishing phase, feed intake was lower in groups fed diets with 200 ppb chromium than in other treatment (p < 0.1). Feed conversion was improved in the groups fed diets with chromium compared with control. Carcass weight was similar among treatments while carcass length was longer in groups fed diets with 200 ppb chromium (p < 0.05). Thinner carcass fat was found with groups fed diets with chromium compared to control. Three A grade of carcasses were from groups fed chromium compared to control. No significant differences were observed with blood glucose, insulin, total cholesterol, triglycerides and non-esterified fatty acid at 60kg body weight. While, at 100 kg body weight, blood triglyceride was lower in groups with 200 and 400 ppb chromium but higher in groups with 100 ppb chromium (p < 0.05). In vitro lipolysis and protein synthesis in adipose tissues were increased as dietary chromium was increased from 0 to 200 ppb (p < 0.1). As a result, 200 ppb chromium in a growing-finishing diet could improve feed efficiency and carcass traits; an increase to 400 ppb has no further effect.
Background: Promoter hypermethylation mediated gene silencing of tumor suppressor genes is considered as most frequent mechanism than genetic aberrations such as mutations in the development of cancers. BRD7 is a single bromodomain containing protein that functions as a subunit of SWI/SNF chromatin-remodeling complex to regulate transcription. It also interacts with the well know tumor suppressor protein p53 to trans-activate genes involved in cell cycle arrest. Loss of expression of BRD7 has been observed in breast cancers and nasopharyngeal carcinomas due to promoter hypermethylation. However, the genetic status of BRD7 in oral squamous cell carcinomas (OSCCs) is not known, although OSCC is one of the most common among all reported cancers in the Indian population. Hence, in the present study we investigated OSCC samples to determine the occurrence of hypermethylation in the promoter region of BRD7 and understand its prevalence. Materials and Methods: Genomic DNA extracted from biopsy tissues of twenty three oral squamous cell carcinomas were digested with methylation sensitive HpaII type2 restriction enzyme that recognizes and cuts unmethylated CCGG motifs. The digested DNA samples were amplified with primers flanking the CCGG motifs in promoter region of BRD7 gene. The PCR amplified products were analyzed by agarose gel electrophoresis along with undigested amplification control. Results: Methylation sensitive enzyme technique identified methylation of BRD7 promoter region seventeen out of twenty three (74%) well differentiated oral squamous cell carcinoma samples. Conclusions: The identification of BRD7 promoter hypermethylation in 74% of well differentiated oral squamous cell carcinomas indicates that the methylation dependent silencing of BRD7 gene is a frequent event in carcinogenesis. To the best of our knowledge, the present study is the first to report the occurrence of BRD7and its high prevalence in oral squamous cell carcinomas.
Purpose: To investigate the expression of Ki67 protein in papillary thyroid microcarcinoma(PTMC), and to analyze its clinical significance. Materials and Methods: Ki67 protein expression was evaluated in the tissues of 108 human PTMC and 50 other benign papillary hyperplasia of thyroid specimens using immunohistochemistry. Results: The expression intensity of Ki67 in PTMC and benign papillary hyperplasia of thyroid specimens were $1.45{\pm}1.83%$ and $0.46{\pm}0.46%$.The positive expression rates were 46.3% and 14%. There were significant differences between these two groups (p<0.01). There was no significant variation of the expression intensity and positive expression rates of Ki67 in PTMC with gender, age, position of the tumor and the level of TSH pre-operation (p>0.05), but these parameters varied with tumor size, invasion by membrane and cervical lymph node metastasis (p<0.05 or p<0.01). Conclusions: The expression of Ki67 in PTMC was related to tumor size, invasion by membrane and cervical lymph node metastasis, and could be the important indicator for judging clinical progress and estimating prognosis.
Kang, Hye Kyeong;Park, Ji Ae;Seo, Kang Seok;Kim, Sang Hoon;Choi, Yun Jai;Moon, Yang Soo
Asian-Australasian Journal of Animal Sciences
/
v.19
no.3
/
pp.329-334
/
2006
Adipocyte-specific secretory factor (ADSF)/resistin, a hormone, is a small cysteine-rich protein secreted from adipose tissue and has been implicated in modulating adipogenesis in humans and rodents. The objective of this study was to clone a gene encoding ADSF/resistin and to characterize its function in Korean Native Cattle (Hanwoo). The coding sequence was 330 base pairs and it encoded a protein of 109 amino acids. An NCBI BLAST-search revealed the cloned cDNA fragment shared significant homology (82%) with the cDNA encoding the human ADSF/resistin. The nucleotide sequence homology of the Hanwoo sequence was 73% and 64% for the rat and mouse, respectively. A 654 bp ADSF/resistin gene promoter was cloned and putative binding sites of transcription factors were identified. Tissue distribution of ADSF mRNA was examined in liver, skeletal muscles (tenderloin, biceps femoris), subcutaneous fat, and perirenal fat by RT-PCR. ADSF mRNAs were detected in fat tissues but not in liver and muscles, suggesting that ADSF/resistin expression may be induced during adipogenesis. Although, the physiological function of ADSF/resistin in the cow remains to be determined, these data indicate ADSF is related to the adipocyte phenotype and may have a possibly regulatory role in adipocyte function.
Chang, Mira;Lee, Jeong-Woong;Koo, Deog-Bon;Shin, Sang Tae;Han, Yong-Mahn
Asian-Australasian Journal of Animal Sciences
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v.23
no.6
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pp.806-813
/
2010
Animal bioreactors have been regarded as alternative tools for the production of limited human therapeutic proteins. The mammary glands of cattle are optimal tissues to produce therapeutic proteins that cannot be produced in large amounts in traditional systems based on microorganisms and eukaryotic cells. In this study, two knock-in vectors, pBCTPOKI-6 and pBCTPOKI-10, which target the hTPO gene on the bovine beta-casein locus, were designed to develop cloned transgenic cattle. The pBCTPOKI-6 and pBCTPOKI-10 vectors expressed hTPO protein in culture medium at a concentration of 774 pg/ml and 1,867 pg/ml, respectively. Successfully, two targeted cell clones were obtained from the bovine fibroblasts transfected with the pBCTPOKI-6 vector. Cloned embryos reconstructed with the targeted nuclei showed a lower in vitro developmental competence than those with the wild-type nuclei. After transfer of the cloned embryos into recipients, 7 pregnancies were detected at 40 to 60 days of gestation, but failed to develop to term. The results are the first trial for targeting of a human gene on the bovine milk protein gene locus, providing the potential for a large-scale production of therapeutic proteins in the animal bioreactor system.
Hong, Young Mi;Lee, Hyeryon;Cho, Min-Sun;Kim, Kwan Chang
Clinical and Experimental Pediatrics
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v.60
no.11
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pp.365-372
/
2017
Purpose: The mechanism for the pathogenesis of adriamycin (ADR)-induced cardiomyopathy is not yet known. Different hypotheses include the production of free radicals, an interaction between ADR and nuclear components, and a disruption in cardiac-specific gene expression. Apoptosis has also been proposed as being involved in cardiac dysfunction. The purpose of this study was to determine if apoptosis might play a role in ADR-induced cardiomyopathy. Methods: Male Sprague-Dawley rats were separated into 2 groups: the control group (C group) and the experimental group (ADR 5 mg/wk for 3 weeks through intraperitoneal injections; A group). Echocardiographic images were obtained at week 3. Changes in caspase-3, B-cell leukemia/lymphoma (Bcl)-2, Bcl-2-associated X (Bax), interleukin (IL)-6, tumor necrosis $factor-{\alpha}$, brain natriuretic peptide (BNP), troponin I, collagen 1, and collagen 3 protein expression from the left ventricle tissues of C and A group rats were determined by Western blot. Results: Ascites and heart failure as well as left ventricular hypertrophy were noted in the A group. Ejection fraction and shortening fraction were significantly lower in the A group by echocardiography. The expression of caspase-3, Bax, IL-6, BNP, collagen 1, and collagen 3 were significantly higher in the A group as compared with the C group. Protein expression of Bcl-2 decreased significantly in the A group compared with the C group. Conclusion: ADR induced an upregulation of caspase-3, Bax, IL-6, and collagen, as well as a depression in Bcl-2. Thus, apoptosis and fibrosis may play an important role in ADR-induced cardiomyopathy.
The fractional synthesis rates(FSR) were measured with 2l-wk and 3l-wk-old broiler breeder pullets and hens to investigate the effect of sexual maturity on FSR. The FSR were obtained from chicken tissues and blood samples using High-Performance Liquid Chromatography/Mass Spectrometry(HPLC/MS). A L-l-13C, 15N -leucine saline solution was infused by bolus injection as a tracer into broiler breeder pullets in the experiment. A rapid HPLC/MS method was developed to measure the isotopic enrichments of leucine in plasma, tissue samples, and eggs. The enrichments of stable isotope leucine incorporated into protein and the enrichments of the stable isotope free leucine were measured in liver, breast muscle and blood samples. Two sets of experiments were conducted. In experiment one, 2l-wk-old, sexually immature broiler breeder pullets were divided into groups of three and blood samples were collected at 20 or 30 min intervals until 1.5 h from initial injection. The pullets were sacrificed in groups of three at varying time intervals for 7 h after injection. The liver, breast muscle and blood samples were removed for analysis. The FSR were estimated to be 8.7l%/day for liver, 4.06%/day for breast muscle, and 5.08%/day for blood samples in 30 minutes after injection from the enrichment ratios. In experiment two, sexually matured 3l-wk-old broiler breeder hens were assorted into groups of three and blood samples were obtained at 20 or 30 min intervals for 2 h. The FSR for blood samples were determined. The broiler breeder hens were sacrificed in groups of three at various time intervals until 7 h after injection and liver, breast muscle and blood samples were removed for analysis. The FSR were calculated to be 5.96%/day for liver. Eggs were collected from five chickens daily for 10 days after large bolus injection. The average of total enrichments of stable isotope in egg albumin was increased by 0.064% at 4 days after injection and was back to normal in 7 days.
Lee, Dae-Weon;Seo, Jong Bok;Kang, Jae Soon;Koh, Sang-Hyun;Lee, Si-Hyeock;Koh, Young Ho
The Plant Pathology Journal
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v.28
no.4
/
pp.409-417
/
2012
We identified two novel expansin (EXP) genes in the expressed sequence tag database of Bursaphelenchus xylophilus, designated as Bx-EXPB2 and -EXPB3. Novel Bx-EXPBs encoded 150 amino acids and their similarities in coding sequence were 70.7-84.0% to the previously reported EXPB1 of B. xylophilus. Bx-EXPB2 and Bx-EXPB3 were clustered with Bx-EXPB1 and Bm-EXPB1, respectively, forming the independent phylogeny with other nematode EXPs. All identified Bx-EXPBs contained the signal peptide and were only expressed during the propagative stage, suggesting that they are secreted to facilitate nematode migration through hosts by loosening cell walls during infection. Quantitative real-time PCR analysis showed that the relative accumulation of Bx-EXPB3 mRNAs was the highest among the three Bx-EXPs examined and the order of mRNA accumulation was as follows: Bx-EXPB3 > Bx-EXPB2 >> Bx-EXPB1. Homology modeling of Bx-EXPBs showed that the structurally optimum template was EXLX1 protein of Bacillus subtilis, whichshared residues essential for catalytic activity with Bx-EXPB1 and Bx-EXPB2 except for Bx-EXPB3. Taken together, Bx-EXPB1 and Bx-EXPB2 may be involved migration through plant tissues and play a role in pathogenesis.
To identify genes involved in rice Pi5-mediated disease resistance to Magnaporthe oryzae, we compared the proteomes of the RIL260 rice strain carrying the Pi5 resistance gene with its susceptible mutants M5465 and M7023. Proteins were extracted from the leaf tissues of both RIL260 and the mutant lines at 0, 24, and 48 h after M. oryzae inoculation and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis identified eight proteins that were differently expressed between the resistant and susceptible plants (three down- and five up-regulated proteins in the mutants). The down-regulated proteins included a triosephosphate isomerase (spot no. 2210), a 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (no. 3611), and an unknown protein (no. 4505). In addition, the five up-regulated proteins in the mutants were predicted to be a fructokinase I (no. 313), a glutathione S-transferase (no. 2310), an atpB of chloroplast ATP synthase (no. 3616), an aminopeptidase N (no. 3724), and an unknown protein (no. 308). These results suggest that proteomic analysis of rice susceptible mutants is a useful method for identifying novel proteins involved in resistance to the M. oryzae pathogen.
Journal of The Korean Society of Grassland and Forage Science
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v.21
no.2
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pp.81-88
/
2001
To investigate C and N metabolisms in response to potassium-deficient stress during regrowth of Italian ryegrass(Lolium multiflorum L.), C and N metabolites were analyzed at day 0 (cutting date), 6, 12 and 24 days after defoliation. K-sufficient (control, +K) and K-absent (-K) nutrition solutions were applied from 7 days before defoliation, and continued for one cycle of 24 days-regrowth period. During 24 days of regrowth dry matter of regrowing shoots and remaining tissues were not significantly different between +K and -K treatment. In remaining stubble, all C compounds in both +K and -K treatment largely decreased (69% to 84% of the initial level) during the first 6 days of regrowth, and then rapidly recovered. The decline of soluble sugars and fructan in roots for the first 6 days much less in the -K medium. Amino acids, soluble and insoluble proteins in stubble also feel down during the first 6 days, thereafter actively replenished in both +K and -K treatment. The decline of nitrate in stubble prolonged to 12 days of regrowth. Initial amounts of all N compounds in roots were significantly lower in the -K medium. Higher accumulation of amino acids and soluble protein in roots in the -K medium was observed after 12 days of regrowth. In regrowing shoots, 3 all carbohydrates increased with a very similar pattern for both treatments. Nitrate was not significantly different between two treatments. Depress of soluble protein accumulation in -K medium was noteworthy after 12 days of regrowth. These results indicated that an active utilization of organic reserves occurred to support regrowth even under K deficient condition with a similar extent with K sufficient condition.
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