This study was conducted to derive the level of importance of HACCP evaluation items in meat packing plants. Based on the results, we proposed a method to increase the objectivity of HACCP criteria for meat packaging plants by giving different evaluation scores to each evaluation item. To assign a differential score for each HACCP evaluation item, the severity of the hazard and the rate of non-compliance in the past 3 years (3,905 of the prerequisite section, 2,057 of the HACCP management section) were analyzed. The results were as follows. The importance and score differentiation of each HACCP evaluation item was derived. The total scores of the prerequisite section and HACCP management section were set at 200 and 100, respectively. However, to maintain objectivity of the HACCP evaluation items, further study is needed to develop detailed criteria for each HACCP evaluation item and score.

Due to their anatomical, physiological and genetic similarities, pig is attractive animal model in biomedical research. In the recent stem cell research era, porcine derived stem cells also gain attention due to its use for the preclinical application of human. Mesenchymal stem cells (MSCs) have been studied by many researchers over decade, and their prospect for clinical application is recognized. Although porcine derived MSCs (pMSCs) have confirmed to be differentiated into various types of cells, such as osteocyte, chondrocyte, neuronal cell, cardiomyocyte and pancreatic β cell, few report has been studied regarding hepatocyte differentiation in vitro. The present study was therefore aimed for bone marrow MSCs derived from pig femur to differentiate into hepatocyte. The cells were confirmed as MSCs by characterizing their morphology, lineage differentiation capacity and surface phenotype. They showed spindle like morphology and adipocytic, osteoblastic, and chondrocytic differentiation potentials and displayed positive expression of mesenchymal markers CD29, CD44 and CD90 while lacked the expression of hematopoietic marker CD45. Under appropriate differentiation conditions, MSCs displayed hepatocyte-like morphology depending on duration of differentiation. The differentiated MSCs into hepatocyte expressed hepatocyte-specific genes including hepatocyte nuclear factor 4 (HNF4), albumin (ALB), alpha fetoprotein (AFP), alpha-1-anti trypsin (A1AT). They also showed hepatocyte-like function, glycogen storage which is identified by PAS staining. Taken together, it concluded that the bone marrow MSCs have the potential to differentiate into hepatocyte. Further studies are needed on additional hepatocytic functional assays, such as low density lipoprotein (LDL) uptake and urea synthesis of differentiated MSC.

The structural diversity and localization of cell surface glycosphingolipids (GSLs), including gangliosides, in glycolipid-enriched microdomains (GEMs) render them ideally suited to play important roles in mediating cell recognition, adhesion, interactions, receptor function, and signaling. Gangliosides, sialic acid-containing GSLs, are most abundant in the nerve tissues. The quantity and expression pattern of gangliosides in brain change drastically throughout development and these changes are mainly regulated through stage-specific expression of glycosyltransferase genes. However, roles of gangliosides in neuronal differentiation of mesenchymal stem cells (MSCs) is unclear. We previously demonstrated for the first time that the glycosyltransferase genes during mouse embryogenesis. So, we investigated the effects of ganglioside gene in differentiation of adipose-derived MSCs (AD-MSCs). GM2 and GD3 ganglioside synthease were increased during neuronal differentiation of AD-MSCs. This study showed that the differentiation of neuronal marker was decreased on the first step of ganglioside synthase UDP-glucose ceramide glucosyltransferase(UGCG) and knock downed GM2 sythase (B4GALNT1). The result of suggested that GM2 and GD3 might be important roles in the neural differentiation of mini-pig AD-MSCs. This work was carried out with the funding of the cooperative research Program for Agriculture Science & Technology Development[Project No. PJ00999901], the Rural Development Adiministration, the KRIBB Research Initiative Program[KGM4251622].

Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.

Adipose tissue is the largest energy storage in the body, with the endocrine, paracrine and autocrine function, and they constitute a network regulatory signal, and participate in energy balance and metabolism regulation in adipose tissue. When adipose tissue is excessively accumulated or obesity, inflammatory signaling pathway is activated as the secretion increase of a variety of inflammatory cytokines, then, the body is under the state of chronic inflammation, causing insulin resistance and many metabolic diseases, such as type 2 diabetes, atherosclerosis, cancer and other chronic metabolic disease, and bringing a serious health crisis to humans. And, excessive fat deposition reduces the feed conversion rate, leading to the increase of livestock and poultry production cost and the reduction of meat food quality. Therefore, the regulation of adipogenic differentiation has become an important field in the study of human health and animal production. 1. The source of adipose tissue The formation of adipose tissue is due to the increase of adipose cell number caused by differentiation and the increase of adipose cell volume and adipose accumulation during development. This process is that adipose mescenchymal stem cells (AMSCs) are transformed to preadipocytes in adipogenic environment, and differentiation related specific transcription factors start to express and induce the specific expression of adipose cell genes and terminal differentiation, finally, mature adipose cells are formed after adipose accumulation. Recent studies have suggested that dedifferentiated fat cells (DFATs) may be an important source of adipose tissue. Mature adipose cells can be dedifferentiated to the sub cells (DFATs) with the dividing ability in vitro culture, and DFATs are pluripotent and can be redifferentiated to adipose cells or transdifferentiated to other cell types, such as cartilage cells, bone cells, muscle cells, etc. by induction. This suggests that DFATs are progenitor cells with the stem cell properties, showing the great potential in tissue engineering and regenerative medicine. Research on the mechanism of DFATs redifferentiation and transdifferentiation has an important significance for human health and animal production. 2. Regulation of adipocyte differentiation and transcription The adipocyte differentiation lies in the transcription level regulation, involving the cascade and cooperative effects of multiple transcription factors, among which, the core transcription factor is peroxisome proliferator activated receptors-γ (PPARγ), which specifically expresses in adipose tissue, combines the promoters of downstream genes promoter and induces their expression, such as lipoprotein lipase (LPL), insulin sensitive glucose transporter 4 (GLUT4), fatty acid synthase (FAS), adipose-specific fatty acid binding protein-2 (AP2) and adiponectin, and promotes the differentiation and maturation of adipose cells. 3. Transcription regulation of PPARγ by Kruppel like factors Kruppel like factors (KLFs) are a class of transcription factors with zinc finger structure, which is involved in the regulation of cell proliferation, cell apoptosis, cell differentiation and tumor formation in a variety of animal cell types. Since KLF15 is first proved to have the transcriptional regulation capability of adipose differentiation by Gray, et al. in 2002, other KLFs are also found to be involved in the adipose differentiation regulation. According to the recent studies of KLFs regulation of adipose differentiation, Christopher, et al. in 2009 summarized KLFs transcription regulation network in the PPARγ upstream. This network includes 8 types of KLFs, namely, KLF2-7, KLF11 and KLF15, among which, KLF2, KLF3 and KLF7 are involved in the negative regulation of adipose formation, while KLF4-6, KLF11 and KLF15 positively regulates adipose formation, and they express according to a certain time sequence during adipocyte differentiation. 4. Regulation of adipose differentiation by curcumin Curcumin is a kind of polyphenols extracted from Curcuma longa L.. Curcumin can reduce the mice obesity formation, directly interfere with the preadipocytes differentiation and decrease the adipocyte number and adipose accumulation. Moreover, curcumin plays a role in the early stage of adipocyte differentiation, and inhibit the mitotic proliferation process and the expression levels of PPARγ, C/EBPα, and certain downstream transcription factors. 5. Regulation of AMSCs and DFATs adipogenic differentiation in pigs It is generally believed that pigs are the most suitable animal models for the application of human clinical medicine. Also, pigs are the largest source of human meat food, and one of animals with the most fat content. Therefore, research on the regulation of porcine adipocyte differentiation has an important significance for the establishment of human disease model and the production of low fat and lean meat pigs. This report summarizes the expression patterns of different KLFs and the effect of curcumin on the KLFs and PPARγ expression during the adipogenic differentiation of porcine AMSCs and DFATs in recent years.

Kisspeptin-10 (KP-10) has been reported to act as a tumor metastasis suppressor via its receptor, G protein-coupled receptor 54 (GRP54). The KP-10/GPR54/BMPs signaling pathway plays an important role in embryonic kidney development. However, its function in osteoblast differentiation is unknown. The aim of this study was to confirm the molecular mechanism for the action of KP-10 on osteoblast differentiation. Expression of the Bone morphogenetic protein-2 (BMP2) and osteogenic genes were determined by RT-PCR and real-time PCR analysis in C3H10T1/2 cells. Transient transfection assays were performed to confirm the effects of KP-10 on BMP2-Luc activity. BMP2 and phospho-Smad1/5/9 protein levels were determined by Western blot analysis. Alkaline phosphatase (ALP) staining experiment was performed to evaluate ALP activity. To further confirm the effect of KP-10-induced GPR54, we used GPR54 Knock out (KO) C3H10T1/2 cells. KP-10 significantly increased osteogenic gene such as Runx2, ALP and Dlx5 in C3H10T1/2 cells. The ALP staining levels were also increased by KP-10. Interestingly, BMP2 mRNA, protein expression and promoter activity were also increased by KP-10. However, KP-10-induced BMP2 expressions were not increased in GPR54 KO cells. These results suggest that KP-10 increases BMP2 expression through GPR54. Next, Western blot analysis shown Smad1/5/9 phosphorylation were enhanced in a time-dependent manner by KP-10 treatment. It is well known that BMP2 increased phosphorylation of Smad1/5/9 via BMP2 receptor. In addition, KP-10 increased NFATc4 mRNA levels and NFATc4 overexpression enhance BMP2 mRNA levels. To confirm the KP-10-induced BMP2 action, we used KP-10-treated medium in wild type cells and GPR54 KO cells. The osteogenic genes were not elevated by KP-10-treated medium (GPR54 KO cells) whereas increased expression levels by KP-10 medium (wild type cells). These data indicate that KP-10 induced osteoblast differentiation through NFATc4-mediated BMP2 signaling.

Myeloid differentiation factor 88 (MyD88) is an intracellular adaptor protein involved in Toll pathway. In this study, we monitored the response of 4 key genes of the insect immune system against Beauveria bassiana JEF-007 in Tenebrio molitor using RT-PCR. To better understand the roles of Toll pathway in mealworm immune system, TmGPR and TmMyD88 was knocked down by RNAi silencing. Target gene expressions were decreased at 6 days post-dsRNA injection. Therefore, mealworms were compromised by B. bassiana JEF-007 at 6 days post-dsRNA injection. Silencing of the TmMyD88 and TmGPR resulted in reducing the resistance of the host to fungal infection. However, only dsTmMyD88 showed significant difference with dsEGFP by statistical analysis, which may be due to partial gene knock down of dsGPR. These results indicate that TmMyD88 is required in mealworms for survival against B. bassiana JEF-007.

무당벌레(Harmonia axyridis)는 종내에서 초시색상패턴이 매우 다양하게 존재한다. 본 논문에서는 서로 다른 색상패턴의 무당벌레를 대상 으로 amplified fragment length polymorphism (AFLP)을 실시하여 무당벌레의 초시색상 패턴간 유전형질의 차이를 확인하고자 하였다. 총 28 개의 프라이머 조합으로 실험을 실시한 결과, 총 2,741 개의 밴드가 검출되었다. 그 중 20 개의 밴드(S1-S20)만이 특정 색상패턴에서 나타났다. 이들 가운데 9 개의 밴드를 색상에 연관된 AFLP 후보 지표로 선발하였다. 밴드 가운데 S1과 S2, S20은 Succinea 1, 2 변이형에 공통적으로 나타났으며, S3와 S5는 Conspicua 변이형에 특이적이었다. 또한 S13는 Spectabilis 변이형에, S15와 S18, S19는 Succinea 2 변이형에 특이적이었다. 특정 색상패턴에만 나타나는 9 개의 AFLP 지표들은 cloning을 통해 염기서열 분석을 실시하였고, GenBank를 이용해 다른 염기 서열과 비교를 해보았지만 아무런 상동성도 찾을 수가 없었다. 무당벌레 종 내 유전적 다양성을 평가한 결과, Spectabilis가 Conspicua보다 Succinea 변이형에 높은 유사성을 보였다. 색상에 연관된 AFLP 후보 지표를 기준으로 sequence characterized amplified region (SCAR) 지표로 변환하여 9 개의 AFLP 분자지표들 가운데에서 5 개만이 SCAR 지표로 전환될 수 있었으며, 이를 통해 AFLP 지표가 무당벌레의 색상과 연관되어 있는지 확인할 수 있었다.

The purpose of this paper is to understand the digital differentiation in social members’ information use via digital devices. Though the attention to the digital differentiation becomes far more increasing, there are only few literatures dealing with quantitative approaches about the digital differentiation. The term ‘digital differentiation’ represents the availability of the information user. It is different from ‘digital divide’ of which the main parameter is accessibility to the information. Once accessibility meets a certain level, availability is considered as a more important factor than accessibility when evaluating the progress of ICT(Information and Communication Technology). We present a model that can describe the digital differentiation phenomenon by using the methodology borrowed from the graph theory, inverse optimization and other established research theory related to digital differentiation. We provide some insights to reduce digital differentiations and therefore our analysis can be used as a guideline for policy maker who desires to mitigate digital differentiations.

Pleurotus cornucopiae (PC) mushrooms is found in the field and commonly known in Japan as Tamogidake mushrooms. Recently it has been reported that PC also alleviating the toxicity of heavy metals. However little is known about mechanism of the action of PC on osteoblast differentiation, especially in transcription factor. Inhibitor of DNA binding-1 (Id-1) function has been linked to the proliferation, migration, and senescence of cells, and studies about relationship between Id-1 and biological function. Therefore, this study was aimed to investigate the effect of PC on osteoblast differentiation and expression of Id-1 and Id-2. PC treatment increased ALP, Col 1 and OCN. PC treatment up-regulated the mRNA levels of Id-1 and Id-2 genes. This PC–induced osteoblast differentiation is more effective in lower doses rather than high doses. This study shows that expression of Id-1 and Id-2 was increased in a dose-dependent manner during PC-induced osteoblast differentiation.

In the present study, we evaluated the effect of CGM on osteogenic differentiation of cultured osteoblasts, and determined whether combination treatment with LLLT had synergistic effects on osteogenic differentiation. The results indicated that CGM promoted proliferation, differentiation, and mineralization of osteoblasts at the threshold concentration of 10 μg/ml; whereas, CGM showed cytotoxic properties at concentrations above 100 μg/ml. ALP activity and mineralization were increased at concentrations above 10 μg/ml . CGM in concentrations up to 10 μg/ml also increased the expression of osteoblast-activated factors including type I collagen, BMP-2, RUNX2, and Osterix. The CGM (50 μg/ml) and LLLT (80 mW for 15 sec) combination treatment group showed the highest proliferation levels, ALP activity, and mineralization ratios. The combination treatment also increased the levels of phosphorylated forms of p38, ATF2, PKD, ERK, and JNK. In addition, the osteoblast differentiation factors including type I collagen, BMP-2, RUNX2, and Osterix protein levels were clearly increased in the combination treatment group. These results suggested that the combination treatment of CGM and LLLT has synergistic effects on the differentiation and mineralization of osteoblastic cells.

최근 농촌 인구의 감소 및 고령화현상, 도농소득격차의 심화로 농촌의 부정적 상황이 지속되고 있으며 농업부가가치 및 농가 소득이 정체되는 것을 타파하기 위해 농업 6차 산업에 대한 연구가 필요하게 되었다. 농업 6차산업의 기대효과는 농업의 고부가가치화, 농가소득 증대, 농촌경 제 활성화를 위한 6차 산업화와 농촌산업화 전략이다. 이를 위해 체계적 인 방안을 모색하여 농산물 생산뿐만 아니라 가공, 제조, 체험, 관광, 숙박, 농가레스토랑, 판매 등 모든 분야가 융 ․ 복합적으로 발전할 수 있는 연구를 지속적으로 진행해야 한다. 특히, 오늘날과 같이 고객의 요구가 다양해지고 시장이 세분화 되고 있는 상황에서 농산물도 품질, 기능, 가격 등의 차별화만으로는 더 이상 경쟁력을 확보할 수 없고, 다른 상품과 달리 소비자가 쉽게 구매결정을 하는 저관여 제품이므로, 더욱 더 포장 디자인의 역할이 중요하다고 볼 수 있다. 상품판매를 위한 포장디자인은 농산물 소비량 증대와 농촌경제를 성장시키는 중요한 역할을 하고 있으며 이에 대한 전략 방안이 필요하다. 현재 농산물 포장디자인은 거의 유 사한 형태와 디자인으로 판매되고 있으며 이는 글로벌 경쟁 제품과 비교 했을때 경쟁력이 약하다고 할 수 있다. 국내 농산물 포장디자인과 해외 사례를 비교 분석함으로써 농촌산업의 활성화를 위한 농산물 포장디자 인의 전략을 제안하고자 한다.

This study is aimed to analyze economical meaning and problems on the industrial differentiation of Korean laver industry. Based on the surveyed data, the export value of korean laver has increased over 28 times for last 20 years($10 million to $300 million) and the separation of farming and processing was an important success factor of rapid growth of korean laver industry. However, the result of the survey shows that the farming profit is 534.1 won out of the total price for a bunch of dried laver, 3,566.3 won. So, farming profit counts for just 15 percent of total price. In contrast, the processing profit is 1,143.5 won and it is 32.1 percent of total price. This means that laver farmers are not being guaranteed their profit properly. This phenomenon is occurred due to lower status of first-hand processors(which produce dried laver) to second-hand processors(which produce seasoned laver) due to advanced payment given by second-hand processors. So, fist-hand processors should provide their product in the price which was designated by second-hand processors. Besides, despite of many business risks caused from climate change and environmental pollution, the market price of raw laver has steadily decreased. For sustainable prosperity of korean laver industry, imbalance on korean laver industry concerning profit sharing is need to be changed. In future, self-processing of dried laver in fishery household and enhancing the role of The Fisheries Cooperative Union in laver industry can be considered.

Myeloid differentiation factor 88 (MyD88) is an intracellular adaptor protein involved in Toll signaling pathway. In this study, we monitored the response of 4 key genes of the insect immune system against Beauveria bassiana JEF-007 in Tenebrio molitor using RT-PCR. TmGPR, antimicrobial peptide Tenecin 1 and Tenecin 2 were up-regulated after fungal infection. To better understand the roles of Toll signaling pathway in mealworm immune system, TmGRP and TmMyD88 was knocked down by RNAi silencing. Target gene expressions were decreased at 2 days post-dsRNA injection, and dramatically reduced at 6 days post-dsRNA injection. Therefore, mealworms were compromised by B. bassiana JEF-007 at 6 days post-dsRNA injection. Silencing of the TmMyD88 and TmGRP resulted in reducing the resistance of the host to fungal infection. However, only dsTmMyD88 showed significant difference with dsEGFP by statistical analysis, which may be due to partial gene knock down of dsGRP. These results indicate that TmMyD88 is required in mealworms for survival against B. bassiana JEF-007.

MicroRNA (miRNA, miR) is essential in regulating cell differentiation either by inhibiting mRNA translation or by inducing its degradation. However, the role of miRNA in odontoblastic cell differentiation is still unclear. In this study, we examined the molecular mechanism of miR-27-mediated regulation of odontoblast differentiation in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. The results of the present study demonstrated that the miR-27 expression increases significantly during MDPC-23 odontoblastic cell differentiation. Furthermore, miR-27 up-regulation promotes the differentiation of MDPC-23 cells and accelerates mineralization without cell proliferation. The over-expression of miR-27 significantly increased the expression levels of Wnt1 mRNA and protein. In addition, the results of target gene prediction revealed that Wnt1 mRNA has an miR-27 binding site in its 3’UTR, and is increased by miR-27. These results suggested that miR-27 promotes MDPC-23 odontoblastic cell differentiation by targeting Wnt1 signaling. Therefore, miR-27 is a critical odontoblastic differentiation molecular target for the development of miRNA based therapeutic agents in dental medicine.

Differentiation of Pleurotus eryngii is laborious and time-consuming tasks especially in mycelial status. For development of a method for differentiation of P. eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotypes. PCR with the each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pair-group method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into 2 clusters. Cluster I and II comprised of 4 and 8 cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivar and strains with 21 allele with a PIC value of 0.9090. These results might be useful to provide an efficient method for the identification of P. eryngii cultivars with separate PCR reactions. (This work was supported by a grant from the Gold Seed Project [Supported by a grant from the IPET (213003-04-3-WTI11), MIFAFF, Republic of Korea.]