Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.
Ha, Min-Seong;Cho, Won-Ki;Kim, Ji-Hyeon;Ha, Soo-Min;Lee, Jeong-Ah;Yook, Jang Soo;Kim, Do-Yeon
Journal of the Korean Applied Science and Technology
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v.35
no.3
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pp.956-962
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2018
Childhood obesity causes a higher risk of obesity, premature death and disability in adulthood. In addition, obese children experience an increased risk of respiratory problems, hypertension, cardiovascular disease, insulin resistance and psychological effects. This study aimed to investigate how an exercise intervention affects health-related physical fitness and inflammatory-related blood factors in obese children after. We hypothesized that there would be positive effects on serum levels of insulin-like growth factor-1 (IGF-1), connecting peptide(C-peptide) and resistin, as well as in muscle and cardiovascular-related physical capacities, after an exercise intervention in obese children. Thirty-seven obese children haveperformed health-related fitness tests and provided blood samples for the analysis of changes in circulating biomarkers, both before and after an 8-week exercise intervention, which includes stretching, aerobic exercise, resistance exercise and sports games. The results indicate that exercise training beneficially affects body compositions, especially percentage body fat and muscle mass, without influencing to body weight and height. The results of the physical fitness tests show that muscle and cardiovascular capacity were increased in obese children in response to exercise training. Simultaneously, the exercise training decreased circulating levels of C-peptide, which equated to a "large" effect size. Although there were no significant effects on the levels of IGF-1 and resistin, they show a "small" effect size. Therefore, our findings suggest that the exercise intervention have beneficial effects on body composition and physical fitness levels in obese children, whichmight be associated with the decline in circulating C-peptide.
Castration of male pig produces significant negative effects on skeletal muscle development. The androgen receptor (AR), two splice variants of insulin-like growth factor-I (IGF-I Ea and MGF) and the myostatin gene may play important roles in this process. In the present study, the expression of AR, IGF-I Ea, MGF and myostatin genes in three skeletal muscles, the brachialis, longissimus and semitendinosus, were studied using real-time quantitative RT-PCR. Our experimental design used 14 pairs of male Landrace sire${\times}$Yorkshire dam piglets. The two piglets in each pair were full sibs, one of which was castrated at 21 d of age; the other remained intact. The study group was divided into subgroups of equal size. Animals in the first subgroup were slaughtered at 147 d and those of the second at 210 d of age. Carcass weight and lean meat yield were similar between boars and barrows at 147 d of age (p>0.05), whereas barrows had lower carcass weight and less lean meat yield at 210 d of age (p<0.05). Castration caused down-regulation of AR gene expression at both 147 and 210 d of age (p<0.05). The two splice variants of the IGF-I gene from porcine skeletal muscle were cloned using RT-PCR, and it was found that MGF differs from IGF-I Ea in having a 52-base insert in the last coding exon of the mRNA. Both splice variants were down-regulated by castration only at 210 d of age (p<0.05). No differences in expression of the myostatin gene were observed between boars and barrows at either 147 or 210 d of age (p>0.05). These results suggest that the downregulation of AR, IGF-I Ea and MGF gene expression following castration helps to explain the negative effect of castration on skeletal muscle development.
Yang, Hoon Joo;Song, Yoon Mi;Kim, Ri Youn;Oh, Ji Hye;Cho, Tae Hyung;Kim, In Sook;Hwang, Soon Jung
Maxillofacial Plastic and Reconstructive Surgery
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v.35
no.1
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pp.31-37
/
2013
Purpose: Maxilla and mandible have different patterns of cortical and trabecular bone and different bone mineral densities, even though both are components of the jaw bone. However, cellular differences between maxilla- and mandible derived osteoblasts (OBs) have rarely been studied. We hypothesize that maxilla- and mandible-derived OBs show different responses to $17{\beta}$-estradiol (E2), which is one of the critical factors for bone formation. This study compares skeletal site-specific cell responses between maxilla- and mandible-derived human OBs to E2. Methods: Maxilla- and mandible-derived OBs derived from an identical donor were separately isolated from a total of five normal healthy subjects aged 18~44 years old, cultured with a treatment of 100 nM estrogen. The responses between maxilla- and mandible-derived OBs to E2 were evaluated and compared using cell proliferation, alkaline phosphatase (ALP) activity and gene expression of osteoprotegerin (OPG), ALP, insulin-like growth factor-1 (IGF-1), and estrogen receptor ${\alpha}$ ($ER{\alpha}$). Results: E2 did not have any distinct effects on the proliferation of both types of OBs. Mandible-derived OBs exhibited higher ALP activity than maxilla-derived OBs in the non-treated condition, which was common in all tested individuals. ALP activities of both types of OBs showed a minor increasing tendency with the treatment of E2, even though there was no statistical significance in some specimens. The gene expression of OB by E2 was diverse, depending on the individuals. There was increased expression of OPG, IGF-1, or $ER{\alpha}$ gene in the part of subjects, which was more repeated in maxilla-derived OBs. In particular, OPG or ALP induction by E appeared less frequently in mandible-derived OBs. Conclusion: Current results revealed that E2 affects maxilla- and mandible-derived OBs into facilitating the osteogenic process despite individual differences. Mandible-derived OBs are less sensitive to bone-forming gene expression by E2.
Kim, Ji-Hee;Park, Hyun-Young;Kim, Dae-Won;Byun, Seung-Jae;Moon, Hyo-Jeong;Lee, In;Yang, Chung-Yong
Journal of Physiology & Pathology in Korean Medicine
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v.26
no.3
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pp.399-407
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2012
To evaluate age and gender differences in the relation of cardiovascular risk factors, cognitive impairment, and subclinical carotid atherosclerosis from aged people using by a cross sectional method. Sixty-nine healthy elders living in the community who had not previously undergone carotid ultrasonography were included. We conducted life style surveys, and cognitive function tests including Korean-mini-mental state examination (K-MMSE) and clinical dementia rating-Korean. Various biomarkers from blood were assessed; fasting insulin-like growth factor-1, lipid-profile, high sensitivity C-reactive protein, total homocysteine, glucose, insulin, Homeostasis model assessment (HOMA) for insulin resistance index, vitamin B12, and folate level. Carotid intima-media thickness (C-IMT), and plaques were measured using carotid ultrasonography and aortic ultrasonography, a valid index of atherosclerosis. For the elderly subjects (aged 65-82 years), cognition impairment was more prevalent in females while subclinical atherosclerosis was more prevalentin males. Increased C-IMT has been kept in males, and C-IMT shows increasing trend and the peak at about 80 year-old in females with increasing age. The significant correlations between C-IMT and many vascular risk factors including age, triglyceride, abnormal homocysteinein male, and K-MMSE, insulin, HOMA index and abnormal aortic ultrasonography in female were different in each gender, with the exception of homocysteine (p<0.05). This data suggests that there were differences of age and gender characteristics in terms of subclinical atherosclerosis, cognitive impairment and vascular risk factors in community-living elders. Further larger and longitudinal studies across entire age are required to better understand the effects of risk factors on subclinical atherosclerosis.
Microarray analysis of gene expression has become a powerful approach for exploring the biological effects of drugs, particularly at the stage of toxicology and safety assessment. Acetaminophen (APAP) has been known to induce necrosis in liver, but the molecular mechanism involved has not been fully understood. In this study, we investigated gene expression changes of APAP using microarray technology. APAP was orally administered with a single dose of 50 mg/kg or 500 mg/kg into ICR mice and the animals were sacrificed at 6, 24 and 72 h of APAP administration. Serum biochemical markers for liver toxicity were measured to estimate the maximal toxic time and hepatic gene expression was assessed using high-density oligonucleotide microarrays capable of determining the expression profile of >30,000 well-substantiated mouse genes. Significant alterations in gene expression were noted in the liver of APAP-administered mice. The most notable changes in APAP-administered mice were the expression of genes involved in apoptosis, cell cycle, and calcium signaling pathway, cystein metabolism, glutatione metabolism, and MAPK pathway. The majority of the genes upregulated included insulin-like growth factor binding protein 1, heme oxygenase 1, metallothionein 1, S100 calcium binding protein, caspase 4, and P21. The upregulation of apoptosis and cell cycle-related genes were paralleled to response to APAP. Most of the affected gene expressions were returned to control levels after 72 hr. In conclusion, we identified potential hepatotoxicity makers, and these expressions profiling lead to a better understanding of the molecular basis of APAP-induced hapatotoxicity.
This study was undertaken to investigate possibility of the allogenic type I collagen inducing osteoinduction or osteoconduction at critical sized bone defect in the rabbit. Twenty Newzealand white rabbit, weighted from 2.8 kg to 3.5 kg, were used in this study. The skull was exposed and two bony defects were created with diameter of 10 mm. Group I(n=10), the bony defects was grafted from the other side bone. Group II(n=10), the bony defects was grafted by the allogenic type I collagen with bone morphogenic protein(BMP). Group III(n=10), the bony defects was grafted by the allogenic type I collagen only. Group IV(n=10), the bony defects was lefted with no grafts. The grafted bones and allogenic type I collagen were investigated with radiologic densitometry, histologic analysis and immunohistochemistry after 12 weeks. No major difference was observed in the gross finding between Group I, II, III, but dura mater was exposed in bony defect,the Group IV. The radiologic study demonstrated more bony opacity in the Group I, but the other groups did not demonstrate a significant difference. In the histologic study, grafted bone edge was completely consolidated with original bone in group I and new bone ingrew into the grafted allogenic type I collagen(group II, III),but there is no bone regeneration from the original bony edge in the group IV. The percent of the new bone formation by cross-sectional area was considered statistically significant at a p value of less than 0.05(p<0.05). In the immunohistochemistry study about BMP antibodies, the group IV demonstrated osteogenic activity in front of advancing original bone edge, in which the osteoblast stained strongly for BMP antibodies, but other group does not demonstrated any osteoblastic expression. There was no immunologic rejection. In conclusion, this results do not demonstrate that the allogenic type I collagen is useful for bone substitute, but the characters of the collagen, such as pliability, easy-handling, sponge-like structure, are useful in interpositional bone graft substitutes. The further evaluation of long term results about the resorption, immunologic tissue reaction, response of applied tissue growth factor to the allogenic collagen is needed.
This study identified hepatic differentially expressed genes (DEGs) affecting the marbling of muscle. Most dietary nutrients bypass the liver and produce plasma lipoproteins. These plasma lipoproteins transport free fatty acids to the target tissue, adipose tissue and muscle. We examined hepatic genes differentially expressed in a differential-display reverse transcription-polymerase chain reaction (ddRT-PCR) analysis comparing high- and low-marbled Hanwoo steers. Using 60 arbitrary primers, we found 13 candidate genes that were upregulated and five candidate genes that were downregulated in the livers of high-marbled Hanwoo steers compared to low-marbled individuals. A BLAST search for the 18 DEGs revealed that 14 were well characterized, while four were not annotated. We examined four DEGs: ATP synthase F0, complement component CD, insulin-like growth factor binding protein-3 (IGFBP3) and phosphatidylethanolamine binding protein (PEBP). Of these, only two genes (complement component CD and IGFBP3) were differentially expressed at p<0.05 between the livers of high- and low-marbled individuals. The mean mRNA levels of the PEBP and ATP synthase F0 genes did not differ significantly between the livers of high- and low-marbled individuals. Moreover, these DEGs showed very high inter-individual variation in expression. These informative DEGs were assigned to the bovine chromosome in a BLAST search of MS marker subsets and the bovine genome sequence. Genes related to energy metabolism (ATP synthase F0, ketohexokinase, electron-transfer flavoprotein-ubiquinone oxidoreductase and NADH hydrogenase) were assigned to BTA 1, 11, 17, and 22, respectively. Syntaxin, IGFBP3, decorin, the bax inhibitor gene and the PEBP gene were assigned to BTA 3, 4, 5, 5, and 17, respectively. In this study, the in silico physical maps provided information on the specific location of candidate genes associated with economic traits in cattle.
Kita, K.;Shibata, T.;Aman Yaman, M.;Nagao, K.;Okumura, J.
Asian-Australasian Journal of Animal Sciences
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v.15
no.12
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pp.1760-1764
/
2002
In order to elucidate the physiological function of circulating IGF-I on muscle protein synthesis in the chicken under malnutritional conditions, we administrated recombinant chicken IGF-I using a osmotic mini pump to fasted young chickens and measured the rate of muscle protein synthesis and plasma metabolite. The pumps delivered IGF-I at the rate of $22{\mu}g/d\{300{\mu}g{\cdot}(kg\;body\;weight{\cdot}d)^{-1}\}$. Fractional rate of protein synthesis in the muscle was measured using a large dose injection of L-[$2,6-^3H$]phenylalanine. Constant infusion of chicken IGF-I did not affect plasma glucose level. Significant interaction between dietary treatment and IGF-I infusion was observed in plasma NEFA and total cholesterol concentrations. When chicks were fasted, IGF-I infusion decreased plasma NEFA and total cholesterol concentrations. On the other hand, IGF-I administration did not affect plasma levels of both metabolites. Fasting reduced plasma triglyceride concentration significantly. IGF-I infusion also decreased the level of plasma triglyceride. Plasma IGF-I concentration of young chickens was halved by fasting for 1 d. IGF-I infusion using an osmotic minipump for 1 d increased plasma IGF-I concentration in fasted chicks to the level of fed chicks. Fasting decreased body weight and the loss of body weight was significantly ameliorated by IGF-I infusion. There was a significant interaction between dietary treatment and IGF-I infusion in the fractional rate of breast muscle protein synthesis. There was no effect of IGF-I infusion on muscle protein synthesis in fed chicks. Muscle protein synthesis reduced by fasting was ameliorated by IGF-I infusion, but did not reach to the level of fed control. Muscle weight of fasted chicks infused with IGF-I was similar to fasted birds without IGF-I infusion, which suggests that muscle protein degradation would be increased by IGF-I infusion as well as protein synthesis in fasted chicks.
Recently the web service area is rapidly growing as the next generation IT paradigm because of increase of concern about SOA(Services-Oriented Architecture) and growth of B2B market. Since a service discovery through UDDI(Universal Description, Discovery and Integration) is limited to a functional requirement, it is not considered an effect on frequency of service using and reliability of mutual relation. That is, a quality as nonfunctional aspect of web service is regarded as important factor for a success between consumer and provider. Therefore, the web service selection method with considering the quality is necessary. This paper suggests the agent-based quality broker architecture and selection process which helps to find a service providing the optimum quality that the consumer needs in a position of service consumer. A theory of agent is accepted widely and suitable for proposed system architecture in the circumstance of distributed and heterogeneous environment like web service. In this paper, we considered the QoS and CoS in the evaluation process to solve the problem of existing researches related to the web service selection and used PROMETHEE(Preference Ranking Organization MeTHod for Enrichment Evaluations) as an evaluation method which is most suitable for the web service selection among MCDM approaches. PROMETHEE has advantages that solve the problem that a pair-wise comparison should be performed again when comparative services are added or deleted. This paper suggested a case study with the service composition scenario in order to verify the selection process. In this case study, the decision making problem was described on the basis of evaluated values for qualities from a consumer's point of view and the defined service level.
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