The Net Promotor Score (NPS) is one of the most well-known metrics for measuring customer loyalty. Originally designed by Reichheld (2003), the measure asks participants to rate their likelihood to recommend the brand on a scale of 0-10, after which respondents are placed into a ‘detractors’ group, ‘passive’ group or ‘promotors’ group. While the measure has attracted much attention due to its simplicity and ease of use, there has equally been much criticism of its reliability, nomological validity and how it is connected to business outcomes. Therefore, the current study aims to understand whether the NPS can be used to identify brand advocacy, and secondly, does the NPS work in a care-based, low switching service context. The study included three unique contexts: at home care, residential care and disability care. In total, there were 611 participants, all of which were based in Australia. A questionnaire was developed and administered to each group and included both quantitative and qualitative questions to understand the consumer experience. The findings supported NPS as an effective metric in a care-based, low-switching context for identifying positive customer advocacy. The implication is that the NPS can be used to track organizational performance; and the extended NPS allows organizations to understand and encourage (address) positive (negative) advocacy. In addition, suggestions for an ‘earned advocacy score’ were provided which may offer a more effective way of understanding consumer experience, while providing clearer, more detailed and more actionable data. The current study provides much needed insight for brand and care organizations to understand how the NPS might be used effectively to facilitate better brand outcomes.

The transcription factor POU5F1, also known as OCT4 plays critical roles in maintaining pluripotency during early mammalian embryonic development and in embryonic stem cells. It is important to establish an OCT4 promoter region-based reporter system to study pluripotency. However, there is still a lack of information about the porcine OCT4 upstream region. To improve our understanding of the porcine OCT4 regulatory region, we identified conserved regions in the porcine OCT4 promoter upstream region by sequence-based comparative analysis using various mammalian genome sequences. The similarity of nucleotide sequences in the 5' upstream region was low among mammalian species. However, the OCT4 promoter and four regulatory regions, including distal and proximal enhancer elements, had high similarity. The putative transcription factor binding sites in the Oct4 5' upstream region nucleotide sequences from mice and pigs also differed. Some of these genes are related to pluripotency, and further research will allow us to better understand the differences in species-specific pluripotency. Next, a functional analysis of the porcine OCT4 promoter region was conducted. Luciferase reporter assay results indicated that the porcine OCT4 distal enhancer and proximal enhancer were highly activated in mouse embryonic stem cells and embryonic carcinoma cells, respectively. Similar to OCT4 upstream-based reporter systems derived from other species, the porcine OCT4 upstream region-based reporter constructs showed exclusive expression patterns depending on the state of pluripotency. This work provides basic information about the porcine OCT4 upstream region and various porcine OCT4 fluorescence reporter constructs, which can be applied to study species-specific pluripotency in early embryo development and the establishment of embryonic stem cells in pigs. This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education (NRF-2017R1D1A1B03032256).

To test the muscle cell specific gene expression, we examined the ability of human α-skeletal muscle actin (ACTA) promoter or human myoglobin (hMb) promoter to direct the expression of the GFP gene in both muscle and non-muscle cells, respectively. C2C12 cells, a mouse myoblast cell line, provide a powerful model to study skeletal muscle differentiation in vitro. We intended to use this cell line as a model for skeletal muscle-specific gene expression during myogenic differentiation from myoblast to myotubes. We compared marker gene expression profiles of proliferating and differentiated C2C12 cells using RT-PCR and fluorescent microscopy analysis. Also, we found that the expression of PCK1 gene under the control of ACTA promoter was proportionally increased as C2C12 differentiated into myotube form. PCK1 is involved in the regulation of gluconeogenesis. In previous research, transgenic mice with overexpressing PCK1 in skeletal muscle showed a greatly enhanced level of physical activity, which extends well into old age. This is due, in part, to an increased number of mitochondria and a high concentration of triglyceride in their skeletal muscles. These mice also had very little body fat, despite eating 60% more than controls. We also constructed a mesenchymal stem cell line and fetal fibroblast cell line for the experiments aiming to make transgenic animals in which the PCK1 gene is specifically expressed in muscle tissue. Accumulated knowledge of this approach could be applicable to a variety of related biological areas including transgenic animal research, gene function study, anti-aging study, etc. This work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Export Promotion Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (316002-5).

Recent transcriptome analyses have shown that long non-coding RNAs (ncRNAs) play prevalent roles in transcriptional regulation. We have reported that promoter-associated ncRNAs (pancRNAs) activate the partner gene expression via local epigenetic changes. Here, we identify thousands of genes under the pancRNA-mediated transcriptional regulation in five mammalian species in common. In the mouse, 1) pancRNA-partnered genes show tissue-specific expression pattern, 2) expression of pancRNAs significantly enriched H3K4me3 and H3K27ac marks towards the partner gene expression, 3) H3K4me1 marks the pancRNA-partnered genes regardless of their expression level, and 4) C- or G-skewed motifs were exclusively overrepresented between -200 and -1 bp relative to the transcription start sites of pancRNA-partnered genes. More importantly, the comparative transcriptome analysis among five different mammalian species using a total of 25 counterpart tissues showed that overall pancRNA expression profile exhibited extremely high species-specificity compared to that of mRNA, suggesting that a significant number of pancRNAs contributed to the enhancement of a set of partner genes' expression in a sequence-specific manner. We conclude that the gain and/or loss of gene-activation-associated pancRNA repertories, caused by formation or disorganization of the genomic GC-skewed structure, finely shapes tissue-specific pattern of gene expression according to a given species.

RNA interference (RNAi) has been considered as an alternative strategy to control agricultural pests whereby double-strandedRNA triggers a potent and specific inhibition of its homologous mRNA. Since small double-stranded RNAs are requiredfor various RNAi applications, there is a need for cost-effective methods for producing large quantities of high-qualitydsRNA. Bacillus thuringiensis produces much insecticidal proteins with expression of their encoding genes being drivenby sporulation-dependent promoters. To develop dsRNA mass-production platform utilizing Bt, the pHT1K-EGFP plasmidvector which has cyt1Aa sporulation-dependent promoter was constructed. The transcriptional level of target gene (EGFP)is higher 113 times than Bt reference gene (ssPE). It was applied to protect honeybee from Sacbrood virus, so targetgene was replaced to SBV-vp1. By ingestion of Bt-derived dsRNA to honeybee shows positive effect on SBV suppression.

Agrobacterium tumefaciens-mediated transformation (AtMT) has been widely used for generation of fungal transformants and recently applied to Beauveria bassiana. In this study to comprehend how the AtMT promoter influences on the expression of selection marker (hygromycin B resistance gene; hph), two different Ti-Plasmids were constructed: pCeg (gpdA promoter-based) and pCambia-egfp (CaMV 35S promoter-based). Putative transformants were subjected to the PCR, RT-PCR and qRT-PCR to inspect the T-DNA insertion rate and gene expression level. In conclusion, more than 80% of the colonies succeeded in AtMT transformation and the hph expression level of AtMT/pCeg colonies was higher than that of AtMT/pCambia-egfp colonies. This result can provide useful information on the AtMT of B. bassiana, especially antibiotics susceptibility and promoter-dependant expression level.

Human interferon alpha 2b (hIFNα-2b) is an important immune regulator widely used in clinic, for the treatment of chronic hepatitis, hairy cell leukemia, chronic myelogenous leukemia and multiple myeloma, etc. The clinically used hIFNα-2b is generally produced by E. Coli, which lacks the post-translational O-glycosylation of naturally synthesized protein, and has a short serum half-life. In this study, we report the successful generation of transgenic chickens that produce hIFNα-2b in the egg white using a feline immunodeficiency virus (FIV)-based lentiviral vector. In preliminary in vitro study, the hIFNα-2b gene under the control of CMV promoter expressed as much as 2,650 ng/㎖ in CEF-LNC-hIFNα-2bW cell. A FIV vector packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected underneath the blastoderm of freshly laid chicken eggs (stage X) to produce a hIFNα -2b transgenic chicken. Out of 187 injected eggs, 55 chicks were hatched after 21 days of incubation, and 27 of the G0 hatched chicks expressed the vector-encoded hIFNα-2b gene. The expression of recombinant hIFNα-2b in transgenic chickens constitutes an important step towards low-cost and full biological activity production of this protein drug in bioreactor. This work was supported by the Bio-industry Technology Development Program, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea, and by a grant from the Next-Generation BioGreen 21 Program (No. PJ011178), Rural Development Administration, Republic of Korea.

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy, if it occurs. Progesterone primes the endometrium for possible implantation and inhibits uterine contraction until birth. 20-alpha hydroxysteroid dehydrogenase (20α-HSD; EC.1.1.1.149) enzyme belongs to the family of aldo-keto reductases. 20α-HSD predominantly converts progesterone into its biologically inactive form 20α-hydroxyprogesterone (20α-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In addition, the activity of 20α-HSD during the luteal phase known to be inhibited by prolactin. In this study, we focused on the analysis of transgenic mice expressing EGFP under control of monkey 20α-HSD promotor in mice testis. The protein expression and localization were detected by Western blotting and Immunohistochemical analysis, respectively. 20α-HSD protein was detected at molecular weight of 37-kDa by Western blotting analysis and EGFP was found at 27-kDa in the testis of TG mice. Also EGFP and 20a-HSD protein expression on 1, 2, 4, 6 and 8 weeks after birth were assessed. Both of them were increased the expression level time-dependently. 20α-HSD were strongly expressed in seminiferous tubule from 1 week after birth as seen in Immunohistochemical analysis. However, EGFP was strongly expressed in the seminiferous epithelial cells. Then, we determined the expression of EGFP mRNA in mice testis. Using primers specific for mouse EGFP, mRNA expression levels were analyzed by RT-PCR. The EGFP molecular weights is 400bp, qRT-PCR results using EGFP primer, The Cq value of the ratio decreased as the age increased. On this basis, mRNA were increased the expression level time-dependently. In conclusion, these observations suggest that the 20α-HSD in testis could be play a pivotal role in the spermatogenesis.

The ANK1 (Ankyrin-1) gene, located on the bovine chromosome 27, encodes a structural protein which forms an important component of the cytoskeleton. Ankyrin belongs to a protein family that links membrane proteins to the underlying spectrin-actin cytoskeleton. Many studies on gene expression regulation have revealed that RNA polymerase binds to the ANK1 gene promoter region. The purpose of the present study was to investigate the relationship between the SNP of the ANK1 promoter region and economic traits in Hanwoo cattle. A total of seven SNPs (C-944T, C-733T, C-687G, A-672G, C-307T, A-104G, C-24T), found in 119 animals, were correlated to economic traits. One of these SNPs, A-104G, was reported for the first time in the present study. Three newly discovered haplotypes were not associated with economic traits. Significant (p<0.05) relationships were found between C-944T and carcass weight, backfat thickeness, loin muscle area and between C-733T, A-672G and intramuscular fat. These results suggest that the SNPs of ANK1 gene may be useful molecular markers for selection of meat yield and quality traits in Hanwoo

For a decade, solution-processed functional materials and various printing technologies have attracted increasingly the significant interest in realizing low-cost flexible electronics. In this study, Cu nanoparticles are synthesized via the chemical reduction of Cu ions under inert atmosphere. To prevent interparticle agglomeration and surface oxidation, oleic acid is incorporated as a surface capping molecule and hydrazine is used as a reducing agent. To endow water-compatibility, the surface of synthesized Cu nanoparticles is modified by a mixture of carboxyl-terminated anionic polyelectrolyte and polyoxylethylene oleylamine ether. For reducing the surface tension and the evaporation rate of aqueous Cu nanoparticle inks, the solvent composition of Cu nanoparticle ink is designed as DI water:2-methoxy ethanol:glycerol:ethylene glycol = 50:20:5:25 wt%. The effects of poly(styrene-co-maleic acid) as an adhesion promoter(AP) on rheology of aqueous Cu nanoparticle inks and adhesion of Cu pattern printed on polyimid films are investigated. The 40 wt% aqueous Cu nanoparticle inks with 0.5 wt% of Poly(styrene-co-maleic acid) show the “Newtonian flow” and has a low viscosity under 10 mPa·S, which is applicable to inkjet printing. The Cu patterns with a linewidth of 50~60 μm are successfully fabricated. With the addition of Poly(styrene-co-maleic acid), the adhesion of printed Cu patterns on polyimid films is superior to those of patterns prepared from Poly(styrene-comaleic acid)-free inks. The resistivities of Cu films are measured to be 10~15 μΩ·cm at annealing temperature of 300 ˚C.

울릉 만병초(Rhododendron brachycarpum)를 5월 27일부 터 9월 27일까지 매월 삽목에 따른 삽수의 발근율은 5월 은 63.3%, 6월은 53.3%, 9월은 41.8%로 높았으나, 8월은 19.8%로 낮았다. 발근율이 높은 처리는 근장 및 근폭 의 생장도 양호하였다. 발근촉진제 처리에 따른 삽수의 발근율은 무처리 23.3%에 비하여 IBA 250mg • L−1는 46.7%와 NAA 125mg • L−1는 53.3%로 양호하였다. 차광 및 LED 보광처리에 따른 삽수의 발근율은 대조구 23.3% 에 비하여 LED 보광이 66.7%로 가장 높았고, 30% 차광이 43.3%, 60% 차광은 30.0%이었다. 근장과 근폭도 발근율이 양호한 LED 보광 및 30% 차광처리는 양호하 게 생장하였다.

산초나무종자는 전통적으로 향신료 및 약용으로 널리 이용되어 왔지만 종자의 발아율이 낮고 묘의 대량생산이 어려워 일부 지역에서만 재배가 되고 있다. 본 연구는 산초나무 종자 휴면타파를 위한 전처리방법을 정립하고 발아촉진물질을 처리하여 출현율을 향상시키고자 하였다. 종자 발아적정온도는 25℃로 서 저온보다 고온에서 발아의 시작이 빨랐고 발아율도 높았다. 종자 휴면타파는 노천매장보다 저온·습윤 처리에서 효과가 높았으며, 종자 외부의 과육 부분을 완전히 제거해야 출현율이 향상되었다. 종자외부의 과육을 제거한 후 270일 동안 저온·습윤 처리하면 출현율이 74.7%까지 향상되었다. 휴면타파 처리기간이 길수록 평균출현일수는 단축되었다. 발아촉진물질 GA3, IAA, kinetin, KNO3 처리는 산초종자의 출현율을 향상시키는데 효과가 있었다. 저온·습윤 120일 처리한 종자는 파종 전 kinetin 2.5mM을 침종처리하면 출현율이 72% 까지 향상되었고 휴면타파 처리기간을 단축시킬 수 있었다.

Silkworm transgenesis is now a routine method leading to a satisfactory yield of transformed animals and the reliable expression of transgenes during multiple successive generations. However, the screening of G1 transgenic individuals from numerous progeny has proved to be difficult and time-consuming work. Previously, we characterized the promoter of heat shock protein 70 from Bombyx mori (bHsp70), which is ubiquitously expressed in all tissues and developmental stages. To investigate the utilization of the bHsp70 promoter to screen transgenic individuals, the EGFP marker gene was inserted into the piggyBac vector under the control of the bHsp70 promoter. Mixtures of the donor and helper vectors were micro-injected into 3,060 eggs of bivoltine silkworms (Keomokjam). EGFP fluorescence was observed in 17 broods of transgenic silkworms under a florescence stereomicroscope. Interestingly, this fluorescent marker protein was detected not only in parts of the embryo segments on the seventh day of the G1 embryonic developmental stage but it was also detected in a part of the body of G1 hatched larvae, in the middle silk gland of G1 fifth instar larvae, and in the wings of seven-day-old G1 pupae and G1 moths. Therefore, we suggest that the bHsp70 promoter can be used for the rapid and simple screening of transgenic silkworms.

Silkworm transgenesis is now a routine method leading to a satisfactory yield of transformed animals and the reliable expression of transgenes during multiple successive generations. However, the screening of G1 transgenic individuals from numerous progeny has proved to be difficult and time-consuming work. Previously, we characterized the promoter of heat shock protein 70 from Bombyx mori (bHsp70), which is ubiquitously expressed in all tissues and developmental stages. To investigate the utilization of the bHsp70 promoter to screen transgenic individuals, the EGFP marker gene was inserted into the piggyBac vector under the control of the bHsp70 promoter. Mixtures of the donor and helper vectors were micro-injected into 3,060 eggs of bivoltine silkworms (Keomokjam). EGFP fluorescence was observed in 17 broods of transgenic silkworms under a florescence stereomicroscope. Interestingly, this fluorescent marker protein was detected not only in parts of the embryo segments on the seventh day of the G1 embryonic developmental stage but it was also detected in a part of the body of G1 hatched larvae, in the middle silk gland of G1 fifth instar larvae, and in the wings of seven-day-old G1 pupae and G1 moths. Therefore, we suggest that the bHsp70 promoter can be used for the rapid and simple screening of transgenic silkworms.

Transgenic chickens have been spotlighted as an highly potent bioreactor for their fecundity, short generation time, and eggs associated with mass production of protein. In this study, we generated transgenic chickens exhibiting oviduct specific expression of human growth hormone fused to human transferrin for oral administration. Gene of the modified growth hormone located at downstream ovalbumin promoter (∼3.6 kb) was introduced to stage X blastodermal cell employing retrovirus vector system. Several transgenic chickens were successfully generated. However, genomic analyses showed unexpected deletion within the transgene. The modification of the transgene seemed to occur during germ cell formation because the deletion was detected only from the sperm DNA of the G0 founder animal. There was no evidence of deletion in the somatic cell DNA samples of the same chicken. Consequently, same pattern of the deletion was confirmed in both somatic and germ cells of the G1 progeny.