• Animal Bioscience
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• 제34권10호
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• pp.1590-1599
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• 2021

Objective: This study investigates the expression patterns of toll-like receptors (TLRs) and intracellular mediators in horse muscle cells after exercise, and the relationship between TLRS expression in stressed horse muscle cells and immune cell migration toward them. Methods: The expression patterns of the TLRs (TLR2, TLR4, and TLR8) and downstream signaling pathway-related genes (myeloid differentiation primary response 88 [MYD88]; activating transcription factor 3 [ATF3]) are examined in horse tissues, and horse peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNs) and muscles in response to exercise, using the quantitative reverse transcription-polymerase chain reaction (qPCR). Expressions of chemokine receptor genes, i.e., C-X-C motif chemokine receptor 2 (CXCR2) and C-C motif chemokine receptor 5 (CCR5), are studied in PBMCs and PMNs. A horse muscle cell line is developed by transfecting SV-T antigen into fetal muscle cells, followed by examination of muscle-specific genes. Horse muscle cells are treated with stressors, i.e., cortisol, hydrogen peroxide (H₂O₂), and heat, to mimic stress conditions in vitro, and the expression of TLR4 and TLR8 are examined in stressed muscle cells, in addition to migration activity of PBMCs toward stressed muscle cells. Results: The qPCR revealed that TLR4 message was expressed in cerebrum, cerebellum, thymus, lung, liver, kidney, and muscle, whereas TLR8 expressed in thymus, lung, and kidney, while TLR2 expressed in thymus, lung, and kidney. Expressions of TLRs, i.e., TLR4 and TLR8, and mediators, i.e., MYD88 and ATF3, were upregulated in muscle, PBMCs and PMNs in response to exercise. Expressions of CXCR2 and CCR5 were also upregulated in PBMCs and PMNs after exercise. In the muscle cell line, TLR4 and TLR8 expressions were upregulated when cells were treated with stressors such as cortisol, H₂O₂, and heat. Migration of PBMCs toward stressed muscle cells was increased by exercise and oxidative stresses, and combinations of these. Treatment with methylsulfonylmethane (MSM), an antioxidant on stressed muscle cells, reduced migration of PBMCs toward stressed muscle cells. Conclusion: In this study, we have successfully cultured horse skeletal muscle cells, isolated horse PBMCs, and established an in vitro system for studying stress-related gene expressions and function. Expression of TLR4, TLR8, CXCR2, and CCR5 in horse muscle cells was higher in response to stressors such as cortisol, H₂O₂, and heat, or combinations of these. In addition, migration of PBMCs toward muscle cells was increased when muscle cells were under stress, but inhibition of reactive oxygen species by MSM modulated migratory activity of PBMCs to stressed muscle cells. Further study is necessary to investigate the biological function(s) of the TLR gene family in horse muscle cells.

• 한국어류학회지
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• 제19권1호
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• pp.1-7
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• 2007

조피볼락의 사육 관리 및 생체역학 모델 결정을 위한 기초자료를 수집하기 위해 수온과 체중에 따른 산소 소비율 (oxygen consumption rate, OCR)을 조사하였다. 절식한 조피볼락 소형어 (습중량, $12.9{\pm}2.7g$, 135마리)와 대형어 ($351.1{\pm}9.2g$, 18마리)를 대상으로 세 가지 수온 (15, 20, $25^{\circ}C$)에 따라 유수식 형태의 호흡실을 이용하여 24시간 동안 5분 간격으로(3반복) 산소 소비율을 측정하였다. 수온과 체중 그리고 두 인자의 상호작용 모두가 조피볼락의 산소 소비율에 유의한 영향을 미쳤다 (p<0.001). 수온 상승에 따라 소형어와 대형어의 산소 소비율은 모두 유의적으로 증가하였다 (p<0.001). 15, 20 그리고 $25^{\circ}C$에서의 시간당 평균 산소 소비율은 소형어에서 각각 414.2, 691.5 그리고 $843.8mg\;O_2\;kg^{-1}h^{-1}$였으며, 대형어에서 각각 182.0, 250.7 그리고 $328.2mg\;O_2\;kg^{-1}h^{-1}$이었다. 실험 수온 조건에서 체중 증가에 따라 산소 소비율은 유의하게 감소하였으며 (p<0.001), 대사율에 미치는 체중의 영향은 0.69~0.75 범위의 지수였다. 조피볼락의 산소 소비율에 미치는 수온 (T)과 체중 (W)의 상관관계는 OCR=89.12+28.79T-1.17W로 나타났다. $Q_{10}$ 값은 $15{\sim}20^{\circ}C$, $20{\sim}25^{\circ}C$ 그리고 $15{\sim}25^{\circ}C$에서 소형어와 대형어는 각각 2.79와 1.90, 1.49와 1.71 그리고 2.03과 1.80이었다. 대사에 의한 에너지 손실은 수온 증가와 체중 감소에 따라 증가하였다(p<0.001). 15, 20 그리고 $25^{\circ}C$에서의 호흡 대사로 인한 일간 평균 에너지 손실은 소형어에서 각각 282.9, 472.3 그리고 $576.3kJ\;kg^{-1}d^{-1}$였으며, 대형어에서 각각 124.3, 171.3 그리고 $224.1kJ\;kg^{-1}d^{-1}$이었다.

• Molecules and Cells
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• 제41권5호
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• pp.413-422
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• 2018

Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.

• Asian-Australasian Journal of Animal Sciences
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• 제24권5호
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• pp.636-642
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• 2011

The effect of different rates of ethanol additive on fermentation quality of napiergrass (Pennisetum purpureum) and residual water soluble carbohydrate were studied in the experiment. The addition rate of ethanol was 0%, 1.5%, 2.5%, 3.5%, 4.5% on fresh weight of napiergrass. The laboratory silos were kept in the room, then were opened on 1, 3, 5, 7, 14, 30 days after ensiling and the changes of silage quality were analyzed, respectively. There was a fast and large reduction in pH from the 5th day of ensiling to below 4.2 except for the 4.5% treatment. After five days the pH of silage decreased slowly and the pH of the ethanol additions was lower than the control. Lactic acid content of ethanol treatments increased significantly (p<0.05) from the 5th day of ensiling, reaching the highest value on either the 7th day or 14th day. The ethanol additive inhibited the break down of silage protein and the ammonia nitrogen content of ethanol addition silage was significantly (p<0.05) lower than the control after 30 days of ensiling. Within the initial first day of ensiling the water soluble carbohydrate content declined quickly. The efficiency of water soluble carbohydrate usage was higher in silage with ethanol than in the control. The acetic acid of ethanol treatment was significantly (p<0.05) lower than control on first and 14th day, but there was no significant (p>0.05) difference among the ethanol addition silages. The volatile fatty acids content of silage increased gradually from the first day of ensiling and reached the peak on 14th day or 30th day and the content of ethanol addition treatment was significantly (p<0.05) lower than the control. The experimental results indicated that adding ethanol inhibited the use of protein and water soluble carbohydrate of aerobic bacteria and reduced the silage losses during the early stage of ensiling and thus supplied more fermentation substrate for lactic acid bacteria and improved the fermentation quality of napiergrass.

• 생물환경조절학회지
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• 제17권1호
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• pp.9-13
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• 2008

본 연구는 고추냉이 수경재배시 유리온실내 무차광, 10, 30, 70% 차광률이 고추냉이 생육 및 광합성 특성에 미치는 영향을 조사하였다. 광도가 증가할수록 엽록소 형광특성 ETR은 함께 증가하였으며, 무차광, 10%, 30%차광간의 차이가 없었으나, 70%차광 처리구는 다른 처리구에 비해 현저히 낮게 나타났다. Yield와 qP는 광도 증가와 함께 감소하였으며, 70% 차광 처리구에서 감소폭이 컸다. 광합성과 기공전도도는 10%차광에서 가장 높았으며, 증산율은 무차광 처리구에서 가장 낮게 나타났다. 고추냉이 엽면적과 생장량은 10%차광에서 가장 높았으며, 엽수는 무차광에서 가장 많고 차광률이 증가할수록 감소하였다. 엽장, 엽폭은 무차광, 10-30% 차광간에 유의적인 차이가 없었으나, 엽병장은 무차광에서 유의적으로 감소하였다. 뿌리의 생장은 70% 차광을 제외하고 처리간에 큰 차이가 없었다. 70% 차광에서 고추냉이 잎과 뿌리의 생장이 현저히 감소하였다 고추냉이를 11월에 정식하여 순환식 담액수경 방식으로 5개월간 재배한 결과 시설내의 차광률은 엽중, 엽병중, 엽면적과 광합성 특성 등을 고려할 때 10% 차광($700{\mu}molm^{-2}s^{-1}$)이 잎 생산에 적합하였다.

• Composites Research
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• 제34권5호
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• pp.311-316
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• 2021

본 연구에서는 3 wt.%, 10 wt.% Cr이 함유된 Fe-합금 분말을 기지 금속으로 하여 분말야금공정을 통해 TiC/steel 금속복합재료를 제조하였다. 이후 복합재료의 기계적 물성에 미치는 열처리의 영향을 고찰하고자, 어닐링 열처리 및 칭-템퍼링 열처리를 실시하였다. 인장, 압축 그리고 항절력 시험과 미세조직 분석을 통하여 Cr 함량 및 열처리 조건에 따른 미세조직의 변화와 구조적 강도 변화의 연관성을 고찰하였다. 10 wt.% Cr을 함유한 TiC/steel의 경우, TiC/steel 계면에서 형성된 조대한 Cr carbide의 영향으로 상온 인장강도 및 항절력이 크게 저하하였다. 이에 반해 압축강도에서는 Cr이 3 wt.%, 10 wt.% 함유된 TiC/steel 복합재료 모두 Cr carbide의 유무와 관계없이 어닐링 열처리 시편에 비해 칭-템퍼링 열처리 후 약 4 GPa에 달하는 높은 압축강도를 보였다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권6호
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• pp.899-904
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• 2018

Objective: This experiment studied the effects of first feed intake time post-hatch on growth performance, nutrient apparent metabolic rate and intestinal digestive enzyme activities in broilers. Methods: Two thousand five hundred and twenty LingNan Yellow broilers were randomly allotted to seven treatments with six replicates of 60 each. The only experimental factor was the first feed intake time which was 18, 24, 30, 36, 42, 48, and 54 hours after hatching. The whole experiment lasted for 21 days. Results: During the whole period, the 30 h treatment had the best body weight and average daily gain (p<0.05), followed by the 24 h group performance optimization. Also, the 30 h group was observed to have the best apparent metabolic rate for ether extract (p<0.05) and crude protein (p<0.05) and the highest activities of amylase, lipase and trypsin in small intestine. And the 24 h group was second only to the 30 h group in terms of the above two measures. Conclusion: These results indicated that the appropriate first feeding time of LingNan Yellow broilers was 24 to 30 hours after hatching.

• Asian-Australasian Journal of Animal Sciences
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• 제20권9호
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• pp.1349-1353
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• 2007

Calcineurin is a calmodulin dependent protein that functions as a regulator of muscle cell growth and function. Agents capable of interacting with calcineurin could have important applications in muscle disease treatment as well as in the improvement of livestock production. Calsarcins comprise a family of muscle-specific calcineurin binding proteins which play an important role in modulating the function of calcineurin in muscle cells. Recently, we described the first two members of the calsarcin family (calsarcin-1 and calsarcin-2) in the pig. Here, we characterized the third member of the calsarcin family, calsarcin-3, which is also expressed specifically in skeletal muscle. However, unlike calsarcin-1 and calsarcin-2, the calsarcin-3 mRNA expression in skeletal muscle kept rising throughout the prenatal and postnatal development periods. In addition, radiation hybrid mapping indicated that porcine calsarcin-3 mapped to the distal end of the q arm of pig chromosome 2 (SSC2). A C/T single nucleotide polymorphism site in exon 5 was genotyped using the denaturing high performance liquid chromatography (DHPLC) method and the allele frequencies at this locus were significantly different among breeds.

• 한국축산식품학회지
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• 제30권5호
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• pp.787-794
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• 2010

Calcium plays a role as a signaling molecule in various cellular events. It has been reported that calcium suppresses adipocyte differentiation only in the early phase of adipogenesis. Herein, we demonstrate that treatment of A23187, a mobilizer of intracellular calcium, on day 4 post adipocyte differentiation could still reduce lipid accumulation in differentiating 3T3-L1 cells for 48 h. In addition, luciferase reporter gene and RT-Q-PCR assays demonstrate that A23187 can selectively inhibit transcriptional activities and expression of PPAR$\gamma$ and LXR$\alpha$, suggesting that A23187 may reduce lipid accumulation in the late phase of adipogenesis via downregulation of PPAR$\gamma$ and LXR$\alpha$ expression and transactivation. Moreover, inhibition of HDAC activity by trichostatin A (TSA) partially blocked A23187-mediated downregulation of transcriptional activities of PPAR$\gamma$ and LXR$\alpha$. Together, our data demonstrate that calcium mobilization inhibits expression and transcriptional activities of PPAR$\gamma$ and LXR$\alpha$, resulting in reduced lipid accumulation in differentiating adipocytes, and thus, mobilization of intracellular calcium in adipocytes may serve as a new preventive and therapeutic approach for obesity.

• Asian Pacific Journal of Cancer Prevention
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• 제16권3호
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• pp.991-996
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• 2015

Side population (SP) cells have stem cell-like properties with a capacity for self-renewal and are resistant to chemotherapy and radiotherapy. Therefore the presence of SP cells in human breast cancer probably has prognostic value. Objective: To investigate the characteristics of SP cells and identify the relationship between the SP cells levels and clinico-pathological parameters of the breast tumor and disease-free survival (DFS) in breast cancer patients. Materials and Methods: A total of 122 eligible breast cancer patients were consecutively recruited from January 1, 2006 to December 31, 2007 at Yunnan Tumor Hospital. All eligible subjects received conventional treatment and were followed up for seven years. Predictors of recurrence and/or metastasis and DFS were analyzed using Cox regression analysis. Human breast cancer cells were also obtained from fresh human breast cancer tissue and cultured by the nucleic acid dye Hoechst33342 with Verapami. Flow cytometry (FCM) was employed to isolate the cells of SP and non-SP types. Results: In this study, SP cells were identified using flow cytometric analysis with Hoechst 33342 dye efflux. Adjusted for age, tumor size, lymph nodal status, histological grade, the Cox model showed a higher risk of recurrence and/or metastasis positively associated with the SP cell level (1.75, 1.02-2.98), as well as with axillary lymph node metastasis (2.99, 1.76-5.09), pathology invasiveness type (1.7, 1.14-2.55), and tumor volume doubling time (TVDT) (1.54, 1.01-2.36). Conclusions: The SP cell level is independently associated with tumor progression and clinical outcome after controlling for other pathological factors. The axillary lymph node status, TVDT and the status of non-invasive or invasive tumor independently predict the prognosis of breast cancer.