• Reproductive and Developmental Biology
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• v.31 no.2
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• pp.83-90
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• 2007

Insulin-like growth factor II (IGF2) and H19 genes are mutually imprinted genes which may be responsible for abnormalities in the cloned fetuses and offspring. This study was performed to identify putative differentially methylated regions (DMRs) of porcine H19 locus and to explore its genomic imprinting in in vitro fertilized (IVF) and somatic cell nuclear transferred (SCNT) embryos. Based on mice genomic data, we identified DMRs on H19 and found porcine H19 DMRs that included three CTCF binding sites. Methylation patterns in IVF and SCNT embryos at the 2-, 4-, $8{\sim}16$-cells and blastocyst stages were analyzed by BS (Bisulfite Sequencing)-PCR. The CpGs in CTCF1 was significantly unmethylated in the 2-cell stage IVF embryos. However, the 4- (29.1%) and $8{\sim}16$-cell (68.2%) and blastocyst (48.2%) stages showed higher methylation levels (p<0.01). On the other hand, SCNT embryos were unmethylayted ($0{\sim}2%$) at all stages of development. The CpGs in CTCF2 showed almost unmethylation levels at the 2-,4- and $8{\sim}16$-cell and blastocyst stages of development in both IVF ($0{\sim}14.1%$) and SCNT ($0{\sim}6.4%$) embryos. At all stages of development, CTCF3 was unmethylated in IVF ($0{\sim}17.3%$) and SCNT ($0{\sim}1.2%$) embryos except at the blastocyst stage (54.5%) of IVF embryos. In conclusion, porcine SCNT embryos showed an aberrant methylation pattern comprised to IVF embryos. Therefore, we suggest that the aberrant methylation pattern of H19 loci may be a reason for increased abnormal fetus after embryo transfer of porcine SCNT embryos.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1649-1656
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• 2015

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.

• Food Science of Animal Resources
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• v.32 no.2
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• pp.228-233
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• 2012

This study was conducted to determine the effects of stevia (Stevia rebaudiana bertoni) and charcoal supplementation on growth performance, immune response and carcass characteristics of finishing pigs. A total of 420 pigs (LYD) were randomly allocated into 7 treatments with 3 replications. Dietary treatments were 1) T1 (basal diet), 2) T2 (basal diet+0.3% stevia), 3) T3 (basal diet+0.6% stevia), 4) T4 (basal diet+0.3% charcoal), 5) T5 (basal diet+0.6% charcoal), 6) T6 (basal diet+0.3% stevia+0.3% charcoal) and 7) T7 (basal diet+0.6% stevia+0.6% charcoal). During the experimental period, average daily gain (ADG) was higher in T2 and T6 groups than the other treatments (p<0.05). Feed conversion ratio (FCR) was higher in T6 group compared to the others (p<0.05). There were no significant differences in total cholesterol level and glutamic pyruvic transaminase (GPT) activity of blood among treatments. In glutamic oxaloacetic transaminase (GOT) activity, T3, T5, T6 and T7 groups showed lower values (p<0.05) compared to T1. Insulin-like growth factor-1 concentration was higher in T2 and T6 groups than the others (p<0.05), but there were no significant differences in immunoglobulin G, lymphocyte, eosinophil, basophil and atypical lymph levels among treatments. In neutrophil, T6 showed higher level compared to the others (p<0.05). In the carcass characteristics, T6 showed higher level of a carcass grade compared to the other treatments. However, carcass length did not show any significant difference among treatments. As a result, dietary supplementation of 0.3% stevia and 0.3% charcoal showed higher ADG, higher FCR and better immune response resulting in better growth performance and carcass characteristics of finishing pigs.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1133-1140
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• 2006

Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.491-496
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• 2015

The purpose of this study was to assess the antioxidant, anti-inflammatory, and immuno-enhancing effects of Daebong persimmon (DP) and Bansi (BS) in vivo. Two types of astringent persimmons (DP and BS) were used for this experiment. C57BL/6J mice were assigned to the following groups: 1) lean control, 2) high-fat diet control (HF), 3) A region DP (3% wt/wt) with HF diet (A-DP), 4) B region DP with HF diet (B-DP), 5) C region DP with HF diet (C-DP), 6) D region BS with HF diet (D-BS), and 7) E region BS with HF diet (E-BS). All mice were sacrificed after 4 weeks of treatment, after which blood and tissues were collected. Antioxidant enzyme activities, inflammatory markers, and immune factors were evaluated. DP and BS treatments did not alter food intake or body weight, compared with HF. Administration of B-DP increased catalase activities in serum. Hepatic levels of malondialdehyde, a product of lipid peroxidation, were significantly lower in A-DP mice than in the HF group. A-DP had down-regulatory effects against inflammation induced by high-fat diet feeding, as shown by significant reduction of interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor-${\alpha}$. Additionally, A-DP treatment exerted an immuno-stimulatory effect, as shown by increasing levels of immunoglobulin G. DP treatment improved the level of insulin-like growth factor-1. These results indicate that DP has beneficial health effects on oxidative stress, inflammation, and immunity in vivo.

• The Korean Journal of Food And Nutrition
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• v.28 no.2
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• pp.228-240
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• 2015

This study was conducted to investigate the dietary and other factors affecting bone mineral density (BMD) in older Korean women. A total of 340 women aged 65 to 74 were recruited from the Kugoksoondam area (Kurye, Goksung, Soonchang and Damyang counties), known as the longevity-belt region in Jeonla province, Korea. They were categorized into two groups according to bone status by T-score : a nonosteoporotic group and an osteoporotic group. Demographic characteristics were collected, as well as information on physical measurements, blood tests for biochemical indicators, health status health-related life style, dietary behavior, favorite food groups, nutrient intake and mini nutrition assessment (MNA). The results are as follows: The mean age of 185 nonosteoporotic women was 69.6 years and that of 155 osteoporotic women was 70.9 years (p<0.001). The mean T-score of the nonosteoporotic group was $-1.5mg/cm^3$ and that of theosteoporotic group was $-3.2mg/cm^3$ (p<0.001). Height and body weight in the nonosteoporotic group were significantly higher than in the osteoporotic group (p<0.001, respectively). There was no significant difference in BMI, although the BMI in the nonosteoporotic group was slightly higher. Waist and hip circumferences in the nonosteoporotic group were significantly higher than in the osteoporotic group (p<0.01, respectively), and the mid upper arm and calf circumferences were also significantly higher than in the osteoporotic group (p<0.001, p<0.01, respectively). The 5 m walking ability was significantly superior compared to the osteoporotic group. Serum levels did not show any significant differences between the groups and were within normal range. The serum total protein, albumin and Insulin-like growth factor (IGFs) levels of the nonosteoporotic group were significantly higher than those of the osteoporotic group (p<0.05, p<0.05, p<0.001, respectively). IGF was 104.7 ng/mL for the nonosteoporotic group and 88.1 ng/mL for the osteoporotic group. Physical activity and appetite in the nonosteoporotic group were significantly higher (p<0.01, p<0.05, respectively). The favorite food groups of the nonosteoporotic group comprised more meats and fish than those of the osteoporotic group (p<0.05, respectively). Nutrient intake was not significantly different, with the exception of niacin intake (p<0.05), but the nutrient intake of the nonosteoporotic group was slightly higher than that of the osteoporotic group. The niacin intake of the nonosteoporotic group and the osteoporotic group were 11.4 mgNE and 10.0 mgNE, corresponding to 103.6% and 90.9% of the Korean EAR, respectively. The MNA score of the nonosteoporotic group was significantly more favorable than for the osteoporotic group. In conclusion, it is necessary to maintain adequate body weight and muscle mass. Habitual physical activity may have a beneficial effect on BMD for older women. Dietary factors, such as meat and fish, higher intake of niacin rich foods and nutrient status for older women also appear to have favorable effects on bone mineral density.