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.

Honey bee, Apis mellifera L., have been widely used as a model organism for biological science because of its highly developed sociality, specialized labor division and passive population management. In order to examine the expression patterns of genes putatively involved in social development in honey bee, quantitative real-time PCR (qRT-PCR) that has been widely used to investigate the expression level of target gene can be used in honey bee study. However, the selection and validation of optimal reference genes is a crucial step prior to running qRT-PCR. In the present study, therefore, the seasonal expression stability of five candidate reference genes in the abdomen of forager and nurse was investigated using three programs (geNorm, NormFinder and BestKeeper), and selected reference genes were validated by the normalization of expression level of vg encoding vitellogenin. Although three programs revealed slightly different gene stability values, overall the combination of two genes (rpS18 and gapdh encoding ribosomal protein S18 and glyceraldehyde-3-phosphate dehydrogenase, respectively) was resulted in the most suitable use for normalization of the target gene in forager. However, a single gene, either rpL32 or rpS18 in the nurse or either rpL32, rpS18, or gapdh in the comparison between foragers and nurses, were suggested to be applied for normalization of seasonal and labor-specific gene expression by qRT-PCR.

Honey bee has been widely used as a model insect for biological sciences because of its sociality and specialized labor division. For the investigation of the seasonal and labor-dependent expression patterns of genes putatively involved in its sociality, quantitative real-time PCR (qRT-PCR) can be applied to quantify gene expression level and selection of reliable reference gene(s) for normalization is an accurate step. In this study, using three softwares (geNorm, NormFinder and BestKeeper), we evaluated seasonal expression stabilities of four reference genes that have been widely used for qRT-PCR in forager and nurse heads. Among four candidates, two genes, rpS18 and gapdh, were suggested to be the optimal reference genes for qRT-PCR.

In the present study, we evaluated the effect of glucose-fructose and sucrose supplementation in glycerol-free tris (GFT) on sperm motility, viability, ROS level, apoptosis (BAX and BCL2) and motility (SMCP) related gene expression of dog sperm according to different post-thaw incubation time. The spermatozoa collected from five dogs were resuspended (5×107 cell/ml) with GFT containing 86 mM glucose and 86 mM fructose (GF-GFT) or 100 mM sucrose (S-GFT). The sperm (500 μl) were loaded in straws, cooled for 50 min at 4℃, frozen using liquid nitrogen (LN2) vapor for 20 min and plunged in LN2. The progressive motility, viability, ROS (H2O2) level and mRNA expression of spermatozoa were evaluated according to post-thaw incubation time (0 h, 3 h and 6 h) at 24℃. ROS was assessed using H2DCFDA stain by flow cytometry. The relative abundances of BAX, BCL2 and SMCP were assessed using quantitative real-time polymerase chain reaction (RT-PCR). The motility of spermatozoa cryopreserved in GF-GFT was increased throughout the post-thaw incubation time. The motility of spermatozoa cryopreserved in S-GFT was increased at 3 h of post-thaw incubation. Whereas, the sperm ROS level in GF-GFT group was decreased at 6 h of post-thaw incubation. However, the ROS level in the group S-GFT was gradually increased with the progress of post-thaw incubation period. The post-thaw incubation had no substantial effect on mRNA expression of BAX, BCL2 and SMCP genes of dog spermatozoa in both the GF-GFT and S-GFT groups. These results indicate that GF supplementation in GFT improves the progressive sperm motility during the 6 h of post-thaw incubation with maintaining similar sperm viability and is more efficient in reducing ROS after 3 h of post-thaw incubation. The addition of GF in GFT for the cryopreservation of dog spermatozoa and post-thaw incubation would open an option to achieve more functioning spermatozoa for future assisted reproduction practices.

The objective of the present study was to evaluate the effect of disaccharides supplementation in glycerol-free tris (GFT) on dog sperm cryopreservation with respect to pH adjustment of extender and post-thaw incubation. The spermatozoa collected from five dogs were resuspended (5×107 cell/ml) with GFT containing 100 mM of lactose (L), trehalose (T) or sucrose (S) or pH adjusted (6.85) 100 mM of lactose (LP), trehalose (TP) or sucrose (SP). The sperm (500 μl) were loaded in straws, cooled for 50 min at 4℃, frozen using liquid nitrogen (LN2) vapor for 20 min and plunged in LN2. After thawing at 37℃ for 25 s in a water bath, the spermatozoa were incubated at 24℃ for 30 min. The progressive motility, viability, mitochondrial membrane potential (MMP) and mRNA expression of SMCP gene were then assessed. The MMP was evaluated by combined JC-1 plus PI staining. The relative abundance of SMCP was assessed using quantitative real-time polymerase chain reaction (RT-PCR). Adjustment of pH in GFT extender supplemented with disaccharides did not improve sperrm motility and viability. In general, post-thaw incubation increased the progressive motility of spermatozoa. The sperm motility in the group S was significantly (P<0.05) higher than other groups regardless of post-thaw incubation time. Similarly, the sperm viability in the group S was significantly (P<0.05) higher following post-thaw incubation. The higher sperm motility in the group S was also supported with the significantly (P<0.05) higher live sperm having high MMP. There was no significant difference in mRNA expression of SMCP gene among the experimental groups. These results indicate that cryopreservation of dog sperm in GFT supplemented with S and 30 min post-thaw incubation at 24℃ could provide better freezability of dog spermatozoa with improved motility and higher MMP.

이 연구는 최근 등장한 영화들 가운데 시간적 구조를 영화 형식에 가장 흥미롭게 표현한 <500일의 썸머>와<그랜드 부다페스트 호텔>을 연구 대상으로 선정하여, 이들의 시간 구조가 어떻게 구성되어 있으며 이러한 구성이 어떻게 시각화되었는지 살펴보았다. 이를 통해 ‘표현의 형식’으로서 이야기의 구성이 스토리를 전달하는 도구일 뿐만 아니라 시각적인 ‘표현의 도구’로써 어떻게 사용되어졌으며, 그러한 이야기의 표현 형식이 스토리텔링에서 어떠한 의미를 지니는 것인지에 대하여 고찰하였다. <500일의 썸머>에서 애니메이션으로 구성된 인터타이틀을 통해 시간의 흐름에 대한 정보를 제공하고 각 날짜별로 남자주인공의 심리상태나 연애상황을 인터타이틀의 영상에 표현함으로써 기억이 갖는 임의적인 시간의 배치로 인한 이야기 구성의 비논리성을 보완하는 기능을 하고 있다. <그랜드 부다페스트 호텔>은 다양한 시기의 이야기를 액자식으로 구성하고 있는데, 각 시기의 이야기가 진행될 때 그 당시 사용했던 화면 비율을 사용함으로써 한 편의 영화 내에서 화면의 크기가 각기 달라진다. 즉, 액자식 구성으로 이야기가 진행됨에 따라 1980년대(1.85:1), 1960년대(2.35:1), 1930년대(1.37:1)의 각 시기별로 그 당시 사용했던 화면 비율을 사용함으로써 각 시대의 분위기와 예술적인 감각 등을 재현하였을 뿐만 아니라, 시간의 배열을 독창적으로 시각화하였다. 이들 두 영화에서 시간의 배열을 시각화한 이러한 표현 방식은 현대 영화들에서 보여지는 상투적인 시간의 왜곡 구조에 창의적으로 접근하였다는 점에서 의미가 있다. 이들 영화의 경우 비연대기적 서사방식 그 자체가 새롭다고 할 수는 없지만, 이러한 서사방식을 시각적으로 표현하는 방식의 독창성이 관객에게 신선함과 퍼즐을 풀어내는 것과 같은 쾌감을 준다. 이러한 시간성 표현들은 단지 영화가 말하고자 하는 내용을 전달하는 형식에 그치는 것이 아니라, 그 표현을 통해서 스토리텔링에서 또 다른 의미를 생산하는 역할을 하게 된다.

The game script was more developed by applying complicated game scenarios and development of real-time game produce technique. However, systematical studies on game scenarios are relatively not enough Therefore, the purpose of the study is to reanalyze elements of game scenarios and to study on contents to be treated by game scenarios to apply the pertinent elements to the game script. To achieve the purpose, the study analyzed the existing researches treating development of game scenarios and researches on time-space expressive methods of game scenarios. On the basis of the results of the analysis, the study found necessary expressive methods to be applied jointly to game scenarios and examined their utility plans.

본 연구의 목적은 직시 표현의 올바른 이해와 사용이 성공적인 의사소통에 도움이 된다는 전제로 출발하여 교사와 학습자의 직시 표현 사용 양상을 확인하기 위하여 한국어 교실에서 이루어진 발화 전사 자료를 분석하여 교사와 학습자의 직시 표현 사용 양상을 살펴보는 데 있다. 직시(deixis)란 한 개인이 발화를 할 때 그 발화의 맥락을 이루는 요소들을 직접적인 언어 표현으로 가리키는 현상을 말하며 가리키고 지시하는 데 사용하는 언어적 형태를 ‘직시 표현’이라고 한다. 직시 표현은 표면적인 의미뿐만 아니라 직시 표현이 사용되었을 때의 발화 상황과 맥락을 이해하여야만 최종적으로 가리키는 바가 무엇인지를 명확하게 알고 성공적인 의사소통을 수행할 수 있다. 본고에서는 인칭 직시 표현, 시간 직시 표현, 장소 직시 표현을 중심으로 그 사용 양상을 살펴보았다. 세 가지의 직시 표현 중에서 가장 빈번하게 사용된 것은 인칭 직시 표현으로 42%를 차지하였다. 다음으로 시간 직시 표현이 30%, 장소 직시 표현이 28%순으로 나타났다. 인칭 직시 표현에서 1인칭 복수로 사용되는 ‘우리’가 44%로 가장 높게 나타났다. 시간 직시 표현에서는 ‘지금’이 45%로 가장 높게 나타났으 며 장소 직시 표현에서는 ‘이, 이거, 이것’이 40%, ‘여기’가 33%로 높은 비율을 차지하였다. 한국어 교실에서 교사와 학습자의 발화 분석을 통해 직시 표현의 사용 양상을 확인할 수 있었다. 앞으로 직시 표현과 사회적 직시 표현을 포함한 직시 표현의 사용 양상과 직시 표현의 사용에서 나타나는 의사소통의 장애 요인, 효과적인 직시 표현의 사용 방안 등에 대한 연구와 분석이 필요할 것으로 보인다.

최근 10년 간 기후변화에 의해 일어나는 태풍, 호우 등의 자연재해는 SOC 시설물 관리에 가장 중요한 이슈가 되고 있다. SOC 시설물 관리를 위해 재난·재해에 의해 발생하는 위험성을 줄이고자 이에 관련된 정보는 지속적으로 수집되며, 관련 정보를 통해 만들어지는 Risk Map은 신속한 정보 변화를 모두 포함하는 하여야 한다. 하지만 주기적으로 변화하는 정보의 경우 그 정보의 형태에 따라 대용량이 될 수 있으며, 정보 전송 및 처리 면에서 신속성이 떨어질 수 있다. Risk Map의 경우 정보의 신속성 및 정확성을 요구하기 때문에, 수집 정보 압축 등의 기법 등을 통해 정보 전송의 효율성을 고려해야한다. 또한, 사용자의 요구에 따라 효율적인 압축 및 신속한 정보를 획득할 수 있는 단말기 혹은 플랫폼이 다양할 수 있으므로 이를 고려하여 정보 전송이 가능하도록 해야 한다. 따라서 재난·재해 관련 정보 및 대용량 공간 정보를 효율적으로 결합한 후, 단말기 성능 및 다양성을 고려한 실시간 처리 기법을 개발하여 결합된 정보의 정밀도를 저해하지 않는 자연스러운 처리가 가능한 Risk Map 동적 표현 기술을 개발할 수 있을 것이다. 본 연구는 향후 고용량 정보 처리 되어 정확도가 높은 Risk Map 정보를 신속히 유관기관에 제공하여 의사결정에 지원 할 수 있다.

Previously, the wheat non-specific lipid transfer proteins (TaLTP), members of a small multigene family, appear to show a complex pattern of expression regulation. For further assessment of expression diversity of the TaLTP genes, we have attempted to evaluate their expression profiles of responses to abiotic stresses via the semi-quantitative RT-PCR method. The expression profiles revealed that the TaLTP genes in group A evidenced highly similar (but not identical) responses against abiotic stresses, whereas much differential expression pattern among genes in each group. The four promoters of TaLTP1, TaLTP7, and TaLTP10 of group A and TaLTP3 of group B were fused to a GUS reporter gene and the recombinant genes were introduced into Arabidopsis. The promoters of TaLTP1, TaLTP7 and TaLTP10 of group A, drove strong but various GUS expression in cotyledons, hypocotyls, epidemic and sub-epidemic cells of young shoots and leaves, floral organs as well as siliques. By contrast, the promoter of TaLTP3 just directed trace expression in cotyledons, young emerged leaves and epidemic cells of flower ovaries. The promoter of TaLTP1 directed the expression in root system whereas the promoters of TaLTP1 and TaLTP10 showed some degree of expression during seed development. The expression diversity of TaLTP genes suggests their multiple physiological functions, evidencing subfunctionalization over evolutionary time.

Gene-expression analysis is increasingly important in biological research, with real-time reverse PCR (RTPCR) becoming the method of choice for high-throughput and accurate expression profiling of selected genes. However, this technique requires important preliminary work for standardizing and optimizing the many parameters involved in the analysis. Plant stress studies are more and more based on gene expression. The analysis of gene expression requires sensitive and reproducible measurements for specific mRNA sequence. Several genes are regulated in response to abitoic stresses, such as salinity, and their gene products function in stress response and tolerance. The design of the primers and TaqMan probes for real-time PCR assays were carried out using the Primer ExpressTM software 3.0. The PCR efficiency was estimated through the linear regression of the dilution curve. To understand the expression pattern of various genes under salt stressed condition, we have developed a unique public resource of 9 stress-related genes in poplar. In this study, real-time RT-PCR was used to quantify the transcript level of 10 genes (9 stress-related genes and 1 house keeping gene) that could play a role in adaptation of Populus davidiana. Real-time RT-PCR analyses exhibited different expression ratios of related genes. The data obtained showed that determination of mRNA levels could constitute a new approach to study the stress response of P. davidiana after adaptation during growth in salinity condition.