돌기해삼 Apostichopus japonicus는 주요 양식 대상 무척추동물로서 우리나라 연안 해역에 서 식하고 있다. 본 연구는 방류 방법에 따른 단기간의 생리학적 스트레스 정도를 평가하기 위하 여 heat shock protein 90 (HSP90) 유전자의 발현 변화를 실시간 정량적 중합효소연쇄반응법 으로 조사하였다. 어린 돌기해삼을 비닐봉지에 산소 포장하여 30분간 수송하거나 방류 해역의 간조기에 1시간 공기 중에 노출된 실험군의 HSP90 유전자 발현은 대조군의 HSP90 유전자 발현에 비하여 통계학적으로 유의미하게 증가하였다(수송 후 실험군 p=0.001; 간조기 실험군 p=0.032). 어린 돌기해삼을 방류 후 6시간까지 분석한 결과, 선상에서 씨뿌림 방식으로 방류된 6시간째의 개체 및 호스를 통과하여 수중으로 방류된 2~6시간째의 HSP90 유전자 발현율은 대 조군에 비하여 약간 감소하는 경향을 보였다(씨뿌림 실험군 p=0.069; 호스 방류군 p=0.093). 한 편, 잠수부에 의해 수중에서 방류된 어린 돌기해삼은 방류 후 시간이 경과할수록 HSP90 유전 자 발현율은 증가하는 패턴이 관찰되었다(p=0.061). 이상의 결과는 방류된 어린 돌기해삼의 단기간 스트레스 반응 연구와 효과적인 방류 방법의 개발에 HSP90 유전자 발현이 유용하게 사용될 수 있음을 시사한다.

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.

최근 살진균제는 세계 식량 안보에 없어서는 안될 필수 요소이며, 그 사용량은 증가하고 있다. 살진균제는 직접적 또는 간접적으로 곤충에 영 향을 미쳐 유전자 및 분자 수준의 변화를 일으킨다. 곤충은 다양한 해독 매커니즘을 통해 살진균제를 포함한 농약으로부터 유발되는 활성산소 (ROS) 독성을 제거한다. 본 연구는 살진균제 캡탄의 비치명적 투여량(0.2, 2, and 20 μg/μL)을 주입 후 갈색거저리의 유충에서 해독효소의 mRNA 발현량을 분석했다. 갈색거저리의 전사체 분석을 통해 해독 매커니즘 관련 유전자인 퍼옥시다제(POX), 카탈라제(CAT), 슈퍼옥사이드 디스뮤타제(SOD) 및 글루타티온-S-트랜스퍼라제(GST)를 발굴하였다. 처리 24시간 후 TmPOX5 mRNA가 유의하게 증가한 것으로 나타났다. 처리 3 시간 후 TmSOD4의 mRNA가 유사하게 증가하였다. 또한 2 μg/μL 처리 24시간 후 TmCAT2의 mRNA 가 유의하게 증가하였다. 캡탄 노출 후 TmGST1 및 TmGST3의 mRNA 발현량도 증가하였다. 결론적으로, TmPOX5 및 TmSOD4 유전자는 갈색거저리에서 캡탄 노출에 대한 바이오마커 또는 생체이물 센서로 작용할 수 있음을 시사한다.

Heat shock proteins (HSPs) are highly conserved cellular proteins that contribute to adaptive responses of organisms to a variety of stressors. In response to stressors, cellular levels of HSPs are increased and play critical roles in protein stability, folding and molecular trafficking. The mRNA expression pattern of two well-known heat shock protein transcripts, HSP70 and HSP90 were studied in two tissues of nerve ganglia, cerebral ganglion and pleuropedal ganglion of Pacific abalone (Haliotis discus hannai). It was observed that both HSP70 and HSP90 transcripts were upregulated under heat stress in both ganglion tissues. Expression level of HSP70 was found higher than HSP90 in both ganglia whereas cerebral ganglion showed higher expression than pleuropedal ganglion. The HSP70 and HSP90 showed higher expression at Day-1 after exposed to heat stress, later decreased at Day-3 and Day-7 onwards. The present result suggested that HSP70 and HSP90 synthesize in nerve ganglion tissues and may provide efficient protection from stress.

Quantitative real-time polymerase chain reaction (RTqPCR) is a rapid and precise method of analysis to quantify the level of gene expression and is widely used in the diagnosis of diseases and quantitative analysis of genes. In RT-qPCR analysis, a reference gene (or housekeeping gene) is used for normalization of experimental results. Since this method of analysis detects a small quantity of the product, it is highly sensitive and it is important for the accuracy and reproducibility of the experiment to select a reference gene suitable for gene expression studies. As the expression levels of the reference gene are affected under different conditions, in order to determine the suitability of the housekeeping gene used as the reference gene, it is necessary to verify the expression stability. In the current study, the stability of the expression of 11 housekeeping genes (B2M, SDHA, GAPDH, RPL13, VIM, EEF1A1, HPRT1, GUSB, RPL19, ACTB, and ABL1) was investigated in the tissues of long-tailed chickens (heart, thigh, and breast). Expression stability evaluation was analyzed with four software: BestKeeper, NormFinder, geNorm, and RefFinder. In our study, GAPDH in heart tissue, HPRT1 in thigh tissue, and RPL13 in breast tissue were selected as the most stable reference genes. Evaluation of the expression stability of housekeeping genes can provide important data in gene expression studies by selecting an appropriate reference gene according to various conditions.

오존은 수돗물 정수장에서 이용되는 소독 물질로 미세오염 물질들을 비롯해서 박테리아나 병원성 미생물체를 효과적으로 제거하는 것으로 많은 연구가 보고되어 있다. 본 연 구에서는 실내 사육 중인 붉은 체색을 지닌 Glyptotendipes tokunagai를 대상으로 서로 다른 농도의 오존 노출에 따른 영향을 파악하기 위해 치사율, 체색 변화와 heat shock protein 70 (HSP70) 유전자 발현을 측정하였다. 오존에 노출 된 G. tokunagai에서 농도-시간 의존적으로 치사율 증가가 관찰되었다. 또한 체색 변화는 오존 농도에 따라 붉은색의 체색이 체절마다 엷어지며 탈색되고 경직되는 현상이 보였다. HSP70 유전자 발현은 저농도인 0.2~0.5 ppm에서 노출 10분과 20분에 유의한 수준으로 높게 나타났으나 (P<0.05), 30 분 노출 후에는 발현량이 감소하는 경향을 보였다. 생리적으로 저산소층에 대해 적응능력이 뛰어난 깔따구 경우에도 오존은 매우 강력한 치사 효과를 유발하여 30분 노출 후 경직과 헤모글로빈 파괴로 인한 탈색이 유발되는 것을 보여주었다. 따라서 본 결과는 수돗물 정수장에서 병원성 미생물을 제거하는 데 사용되는 오존이 수생물에 주는 영향성을 파악하는 기초자료로서 활용될 수 있을 것이다.

In this study, we investigated whether infusion of colorectal cancer cell line and PMSG could increase endometrial cancer. As a result, our study confirmed that the injection of colorectal cancer can cause inflammation and cancer in the uterus and increase the VEGF gene in the uterus. The study also found that endometrial cancer was associated with PMSG.

Enamel knot (EK)—a signaling center—refers to a transient morphological structure comprising epithelial tissue. EK is believed to regulate tooth development in early organogenesis without its own cellular alterations, including proliferation and differentiation. EKs show a very simple but conserved structure and share functions with teeth of recently evolved vertebrates, suggesting conserved signaling in certain organs, such as functional teeth, through the course of evolution. In this study, we examined the expression patterns of key EK-specific genes including Dusp26 , Fat4, Meis2, Sln , and Zpld1 during mice embryogenesis. Expression patterns of these genes may reveal putative differentiation mechanisms underlying tooth morphogenesis.

The oral care probiotic strain Weissella cibaria CMU (oraCMU) inhibits volatile sulphur compounds associated with halitosis, presumably by inhibiting the growth of associated oral pathogens. In the present study, we investigated whether oraCMU inhibits the production of these compounds by suppressing the expression of mgl . This gene encodes L-methionine-α-deamino-γ-mercaptomethane-lyase (METase) and is involved in the production of methyl mercaptan (CH3SH) by Porphyromonas gingivalis . Therefore, we specifically investigated the effects of oraCMU on the growth, CH3SH production, METase activity, and mgl expression of P. gingivalis . The minimum inhibitory concentrations of cell-free supernatant and secreted proteins from oraCMU were 125 mg/mL and 800 µg/mL, respectively. At sub-minimum inhibitory concentration levels, these metabolites inhibited CH3SH production, but they also reduced P. gingivalis viability. Only heat-killed oraCMU decreased CH3SH production without affecting P. gingivalis viability. Heat-killed oraCMU also inhibited METase activity toward L-methionine and mgl mRNA expression (p < 0.05). In summary, we demonstrated the inhibition of volatile sulphur compounds via the antimicrobial action of oraCMU and, for the first time, the inhibition of such compounds by heat-killed oraCMU, which occurred at the molecular level.

Hypoxia is one of the most common features of cancer. It is also associated with cancer progression and the acquisition of aggressiveness, which includes invasion and metastasis. Oral squamous cell carcinoma accounts for 90% of all oral cancers, and its 5-year survival rate is about 50%. Despite various attempts and trials, its prognosis has not improved. Among numerous adverse prognostic factors, hypoxia is suspected as one of the most important factors, as it increases the aggressiveness of oral cancer cells. We attempted to observe the effect of hypoxia on the expression of epithelial-mesenchymal transition markers in oral cancer cells. We analyzed and compared both the mRNA and protein expression levels of epithelial-mesenchymal markers using qRT-PCR and western blotting in both normoxic and hypoxic YD10B oral squamous cell carcinoma cells. Eighty-six genes were analyzed through real-time PCR using commercial microarray plates, performed in triplicate. Among the 86 genes, the expression of 24 were increased (≥ 2 fold) by hypoxia, while that of three genes was decreased (≥ 2 fold). Hypoxia significantly affects epithelial-mesenchymal transition-related genes. Further studies on the regulation of these genes may help to develop more efficient therapeutic modalities for oral cancer and to improve prognosis of oral cancer patients.