The prevalence of cancer in companion dogs is growing nowadays with the increasing worldwide population of domestic dogs. Since there is a less established standard of care in veterinary medicine, investigational treatments, such as the development of biomarkers can be considered as a therapeutic intervention for early diagnosis. Despite the enormous efforts that have been invested in the search of biomarkers, still, there is a need for easy detection of significant biological markers for predicting canine cancers at an early stage. In this study, we have analyzed the expression pattern of previously reported 46 canine cancer-associated candidate genes in blood specimens using real-time qPCR. We hypothesized that analysis of gene expression in blood would provide preliminary evidence of local or systemic immunogenic response which further contribute to the easy and early diagnosis of canine cancer from blood specimen as an analytical tool. The datasets included a total of 22 blood samples collected from different breeds of dogs diagnosed with cancer and five from healthy normal dogs. RT-qPCR analysis was performed by employing the SYBR Green PCR mix to assess the expression of these 46 genes in a total of 27 samples. From our result, a total of nine genes (ROS1, C1QA, CD48, IL1b, TLR2, IL2R, CHI3L1, CTSS, and TLR7) were found to be significantly up-regulated (p < 0.05 and p < 0.01) in the cancer samples compared to non-cancer samples. The relative expression level of ROS1, C1QA, CD48, IL1b, TLR2, IL2R, CHI3L1, CTSS, and TLR7 genes was 5.74, 4.78, 3.94, 2.94, 2.57, 2.53, 2.50, 2.04, and 2.57, respectively, in cancer samples compared to non-cancer samples. Thus, our results reveal several highly expressed cancer genes that can be therapeutic target genes for further testing in canine cancers.

Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP × Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (|log2FC| > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

Cluster of differentiation (CD) 24 or heat stable antigen 24 (HSA) molecule is a mucin-type glycoprotein attached to the cell surface by glycosylphosphatidylinositol (GPI)-anchor, promoting adhesive interactions between cells or in extracellular matrix. The aim of this study was to determine the not yet fully identified porcine CD24 gene and protein structure using computational analysis and to validate variants reported in exons of CD24 gene using direct sequencing. A total of 59 samples belonging to Yorkshire, Landrace, Berkshire, Jeju black pig and wild boar were used in the study. Human CD24 mRNA sequences were used as a reference and subjected to BLAST searches to retrieve the orthologous expressed sequence tags (ESTs) or cDNA sequences against NCBI and Ensemble databases. Assembled ESTs and retrieved cDNA sequences for the porcine CD24 gene were used for specific BLAST search to determine its genomic structure. We found porcine CD24 gene to consist of two exons and a relatively long intron. Second exon of porcine CD24 gene had a long 3’ untranslated region (UTR) and was very similar to that of human, mouse, rat, and sheep. The sequence homology of porcine CD24 protein was 65.38-84.62%, when analyzed with amino acid sequences of rat, mouse, human, cattle, and sheep CD24 protein. N-terminal signal sequence, O-glycosylation sites and GPI-anchoring signal sites were also predicted in pig, which showed these motifs to be evolutionary conserved across the species. Variant analysis in exonic regions of porcine CD24 among the multiple breeds showed that only second exon contained eight SNPs and three insertions in a 3’ UTR. Taken together, this study reports putative porcine CD24 gene and its protein structure using in silico approaches, which will be helpful for any further functional studies.

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor’s structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/ chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

송아지 조기 이유는 반추위 발달을 포함하여, 성장과 고기의 육질 형성에 밀접한 관련이 있다. 이유 전 올바른 영양 관리 및 사료 급여에 대한 기초 기반 연구를 위하여 한우 송아지 이유용 사료 급여에 따른 반추위 발달에 미치는 영향을 조사하였다. 한우 송아지 총 17두를 공시축으로 하여 사료급여 체계에 따라 4개의 그룹으로 나누었으며, 그룹 1은 분유, 농후사료를 급여하였고, 그룹 2는 분유, 농후사료, 조사료, 그룹 3은 분유, 농후사료, 전분, 그룹 4는 관행적인 대조구로 모유를 급여하였다. 그룹별 사료 섭취량은 그룹1, 2, 3 각각 0.9 kg/d, 0.88 kg/d, 0.87 kg/d였고, 그룹2에는 조사료 티모시 0.18kg/d를 추가적으로 급여하였다. 사료 에너지가는 그룹 1, 2, 3 공통적으로 DM 88%, CP 15%, TDN 68% 수준이고, 그룹 3은 전분 30%를 추가 공급하여 TDN 함량을 73%로 높였다. 어미젖을 포유한 후 2주령 후부터 10주령까지 8주간에 걸쳐 각 사료 체계에 따라 급여 후, 각 그룹으로부터 반추위를 적출하여 반추위의 유전자 발현을 분석 하였다. 상피 세포의 성장과 관련이 있는 KRT4 유전자는 그룹 3에서 높은 발현을 보였고, 각종 염증성 질환에 대한 면역 반응과 관련이 있는 BPIFA1 유전자는 그룹 1, 3에서 발현이 증가함을 확인할 수 있었으며, 반추위 발달에 중요한 역할을 하는 ACAT1 유전자는 조사료를 급여한 그룹 2에서 발현율이 높게 나타났다. 결과적으로 송아지 조기 이유 및 양질의 조사료 급여는 반추위 발달에 긍정적으로 작용하며, 이를 실험 결과를 통하여 유전자 발현 수준에서 확인 할 수 있었다.

Muscle satellite cell (SC) is responsible for postnatal muscle growth, repair, and regeneration. Satellite cell is an im-portant source of multi-potent stem cell process and differentiation into adipogenic, myogenic, and osteoblastogenic. The objective of this study was to identify alter of transcriptome during differentiation in porcine satellite cell and to elevated transcriptome at different stages of postnatal development to gain insight into the differences in differ-entiated PSC. We used RNA-seq technique to investigate the transcriptomes during differentiation in pig muscle. Sequence reads were obtained from Illumina HiSeq2000. Differentially expressed genes (DEG) were detected by EdgeR. Gene ontology (GO) terms are powerful tool for unification among representation genes or products. In study of GO biological terms, functional annotation clustering involved in cell cycle, apoptosis, extracellular matrix, phosphoryla- tion, proteolysis, and cell signaling in differences stage. Taken together, these results would be contributed to a better understanding of muscle biology and processes underlying differentiation. Our results suggest that the source of DEGs could be better understanding of the mechanism of muscle differentiation and transdifferentiation.

Satellite cells were derived from muscular tissue in postnatal pig. Satellite cell is an important to growth and development in animal tissues or organs. However, the progress underlying induced differentiation is not clear. The aim of this study was to evaluate the morphologic and the transcriptome changes in porcine satellite cell (PSC) treated with insulin, rosiglitazone, or dexamethasone respectively. PSC was obtained from postnatal muscle tissue. In study 1, for study the effect of insulin and FBS on the differentiated satellite cells, cells were cultured at absence or presence of insulin treated with FBS. Total RNA was extracted for determining the expression levels of myo-genic PAX3, PAX7, Myf5, MyoD, and myogenin genes by real-time PCR. Myogenic genes decreased expression levels of mRNA in treated with insulin. In study 2, in order to clarify the relationship between rosiglitazone and lipid in differentiated satellite cells, we further examined the effect of FBS on lipid accumulation in the presence or absence of the rosiglitazone and lipid. Significant differences were observed between rosiglitazone and lipid by FBS. The mRNA of FABP4 and PPARγ increased in rosiglitazone treatment. In study 3, we examined the effect of dexame-thasone on osteogenic differentiation in PSC. The mRNA was increased osteoblasotgenic ALP and ON genes treated with dexamethasone in 2% FBS. Dexamethasone induces osteoblastogenesis in differentiated PSC. Taken together, in differentiated PSCs, FABP4 and PPARγ increased to rosiglitazone. Whereas, no differences to FBS and lipid. These results were not comparable with previous reports. Our results suggest that adipogenic, myogenic, and osteoblasto-genic could be isolated from porcine skeletal muscle, and identify culture conditions which optimize proliferation and differentiation formation of PSC.

Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90∼100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson’s, oil red O, and Alizarin red staining respectively. We per-formed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator- acti-vated receptor gamma (PPARγ) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteo-blast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were indu-ced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strat-egies for augmenting meat quality.

돼지의 육질과 관련하여 지방침착의 중요도가 높아짐에 따라 지방대사 또는 지방축적과 관련한 후보유전자 발굴 을 위해 등지방 조직 유래의 cDNA microarray를 이용하여 품종별 지방함량관련 조직의 유전자 발현 양상을 분석하 였다. 그 결과 재래돼지의 등지방 조직에서 SCD (stearoyl-CoA desaturase)와 ELOVL6 (elongation of very long chain fatty acid 6)가 높은 발현을 보였고, 간에서 FMO1 (hepatic flavin-containing mono oxygenase)이 높은 발현을 보였으며, 요크셔는 간에서 FGG (fibrinogen gamma polypeptide)와 C3d (com- plement component c3d), 등지방에서 COL3A1 (type Ⅲ collagen alpha 1)이 높은 발현을 보이는 것을 확인하였다. Real-time PCR을 통해 microarray data의 validation과 품종간 유전자 발현량을 비교하여, 위의 유전자들 중 지방함량에 따른 차등발현 유전자로 SCD, ELOVL6 그리고 FGG를 선정하였다. 선정된 유전자 SCD, ELOVL6, FGG는 지방대사에 관여하며 근내지방 함량에도 영향을 미쳐 향후 육질개선에 유용한 후보유전자로 서 활용가능성을 제시하였다.