Characteristics of induced pluripotent stem (iPS) cells are consistent with those of embryonic stem (ES) cells. However, exogenous genes integrated by using retrovirus delivery systems cannot be completely removed from the cells. In a recent report, activation-induced cytosine deaminase (AID) and thymine DNA glycosylase (TDG) can induce pluripotency state in mouse differentiated cells through the process of DNA demethylation. Thus, we hypothesized that the two reprogramming factors may convert efficiently bovine differentiated cells into pluripotency state. So, genes of AID and TDG were integrated into pCMV6-AC-IRES-GFP-Puro expression vector, which was transfected into bovine differentiated cells. As results, the colonies derived from AID+TDG-induced bovine cells were formed on day 7 after culture. The number of AP positively colonies in AID+TDG-induced bovine cells was significantly higher than in AID-induced bovine cells (p<0.05). Additionally, expression of pluripotent genes (OCT-3/4, NANOG, SOX2) was slightly increased in AID+TDG-induced bovine cells, as compared to AID-induced bovine cells. Protein expressions of OCT-3/4, NANOG and SOX2 in AID+TDG-induced bovine cells were slightly increased rather than AID-induced bovine cells. Finally, DNA demethylation in the promoter regions of pluripotent markers in AID+TDG-induced bovine cells was increased than that of AID-induced bovine cells. In conclusion, pluripotent stem cells could be efficiently produced from bovine differentiated cells by using non-integrating delivery system with the reprogramming factors (AID and TDG).
Genotyping-by-sequencing (GBS) is a cost-efficient method which can be useful for SNP marker discovery in a population of interest. GBS is genome reduction sequencing method using restriction enzyme. The quality of DNA is a key factor which could have an influence in downstream analysis. However, there have not been many studies which investigated the impact of DNA degradation and the quality of the data on marker discovery. In this study, GBS data of 6 Hanwoo samples (H1~6) showing differing level of DNA degradation were compared. Re-sequencing pipeline was followed to investigate the impact of DNA degradation on marker discovery. As a result, we found that the quantity and quality of SNPs were not affected in the sample H5 and H6 with moderately degraded DNA. On the other hand, marker discovery was greatly affected in samples with severe DNA degradation (H3 and H4). The findings in this study support that GBS is a robust genotyping method towards moderate DNA degradation.
The objective of this experiment was to determine the effect of supplementation of hot melt extrusion (HME) processed Zn sulfate on growth performance, nutrients digestibility, small intestinal morphology and excretion of Zn in weanling pigs. A total of 200 piglets of mixed sex randomly allotted to four treatments on the basis of initial BW (7.15±0.81 kg). There were five replicates in each treatment with 10 pigs per replicate. The experimental treatments consisted of: 1) basal diet containing ZnSO4; 2) basal diet containing Zn-Methionine (ZnMet); 3) basal diet containing low level of nano-Zn as HME (ZnHME50); 4) basal diet containing medium level of nano-Zn as HME ZnSO4 (Zn-HME75). The average daily gain was improved by the ZnMet and ZnHME75 compared with the pigs fed ZnSO4 supplemented diets (p=0.009). Moreover, ZnHME75 and ZnMet affected on the ATTD of CP during phase 2 (p=0.014). The villus height (VH) was affected by increasing when pigs fed diets supplemented the ZnHME75 (P=0.044). The pigs fed diets supplemented ZnHME50 had significantly the lowest (p=0.037) Zn content in liver compared with other treatments. The Zn content in the feces was significantly higher (p<0.001) in ZnSO4 and ZnMet compared with ZnHME50 or ZnHME75. In conclusion, it could be concluded that dietary Zn can be reduced by 25% with ZnHME without any detrimental effect on performance of weanling pigs.
The present study investigated the effects of dietary supplementation of purified amino acid (PAA) isolated and purified from animal slaughter house blood on growth performance and immune response of weanling piglets. A total of fifteen, 28 days old, weaned pigs were randomly distributed into three treatment groups, 5 pens per treatment (one head per pen). Each experimental diet was provided to five piglets per group (T0, 0% PAA; T1, 0.1% PAA; T2, 0.5% PAA). The feed consumption was calculated daily and body weight on a weekly basis. The blood samples were analyzed for plasma concentration of biochemical parameters and cytokines using ELISA Kit assay. Pigs fed with the 0.5% PAA have greater average daily weight gain (ADG) and feed efficiency (FE) as compared to those of T0 and T1. However, ADG and FE of T0 and T1 have no remarkable differences. The plasma cytokine levels were lower in T2 as compared to T0 and T1. The blood parameters like total bilirubin, blood urea nitrogen (BUN), glutamate oxalate transaminase (GOT) and glutamate pyruvate transaminase (GPT) were within the normal range in all the treatment groups. The study indicated that supplementation of 0.5% PAA have positive effect on the growth performance of the weanling pigs. Moreover, supplementation of the amino acid isolated and purified from animal slaughter house blood has no adverse effect on palatability of the feed and health of the animals.
The objective of this experiment was to investigate amino acids and mineral compositions of the four major muscles (LD: longissimus dorsi, PM: psoas major, SM: semimembranosus, and GM: gluteus medius) from Korean native black goat (KNBG). Five uncastrated male KNBGs of 36 months of age were commercially slaughtered and the four muscles were sampled to determine concentrations of myoglobin, collagen, amino acids and minerals. There were significant differences (p<0.05) in myoglobin and collagen content among the four muscles. Myoglobin content of PM was significantly higher (p<0.05) than those of SM and GM. Collagen content of SM was significantly higher (p<0.05) than that of LD. There were significant differences (p<0.05) in cystine and leucine concentrations among the four muscles, and PM had the highest concentrations of the two amino acids compared to other muscles (p<0.05). Also, there were significant differences (p<0.05) in K, P, Mg and Fe contents among the four muscles from KNBG. PM had the highest mineral content while the lowest mineral content was observed in LD. In addition, Fe contents of PM and GM were higher than those of LD and SM (p<0.05). These results indicated that amino acids and mineral content vary considerably with the anatomical location of muscles of Korean native black goat.
Bropirimine, a class of antineoplastic agents, is known as one of the potent immunomodulators and is currently under clinical development for the treatment of cancer. However, the effect of bropirimine on the cow remains unknown as a therapeutics agent. In this experiment, the effect of bropirimine in the peripheral blood mononuclear cells (PBMCs) stimulated by lipopolysaccharide (LPS) or concanavalin-A (Con-A) was examined. Jugular venous blood was collected from Korean Hanwoo calves and PBMCs were isolated. It was used to study the effect of bropirimine upon stimulation with LPS or Con-A for 72 hours. The expression pro-inflammatory cytokines like Tumor Necrosis Factor α (TNF-α) and Interferon γ (IFN-γ) were confirmed. Bropirimine significantly inhibited LPS- or Con-A-induced TNF-α and Con-A-induced IFN-γ in dose-dependent manner. Furthermore, Bropirimine inhibited TNF-α and Con-A mRNA expression at the transcription level. These results clearly indicated that bropirimine inhibited LPS or Con-A stimulated up-regulation of proinflammatory cytokines in a dose-dependent manner without conspicuous cytotoxicity. The bropirimine has potential to protect cow from LPS or Con-A induced endotoxin shock, possibly through inhibition of the production of proinflammatory cytokines. It suggesting that bropirimine may be a novel therapeutic agent for the prevention of inflammatory diseases. This result revealed specific features of the immune responses depending on the bropirimine compound and would help to knowledge of bovine immunity.