• Animal Bioscience
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• v.37 no.3
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• pp.437-450
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• 2024

Objective: Vanin-1 (VNN1) is a pantetheinase that catalyses the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies have shown that the VNN1 is specifically expressed in chicken liver which negatively regulated by microRNA-122. However, the functions of the VNN1 in lipid metabolism in chicken liver haven't been elucidated. Methods: First, we detected the VNN1 mRNA expression in 4-week chickens which were fasted 24 hours. Next, knocked out VNN1 via CRISPR/Cas9 system in the chicken Leghorn Male Hepatoma cell line. Detected the lipid deposition via oil red staining and analysis the content of triglycerides (TG), low-density lipoprotein-C (LDL-C), and high-density lipoprotein-C (HDL-C) after VNN1 knockout in Leghorn Male Hepatoma cell line. Then we captured various differentially expressed genes (DEGs) between VNN1-modified LMH cells and original LMH cells by RNA-seq. Results: Firstly, fasting-induced expression of VNN1. Meanwhile, we successfully used the CRISPR/Cas9 system to achieve targeted mutations of the VNN1 in the chicken LMH cell line. Moreover, the expression level of VNN1 mRNA in LMH-KO-VNN1 cells decreased compared with that in the wild-type LMH cells (p<0.0001). Compared with control, lipid deposition was decreased after knockout VNN1 via oil red staining, meanwhile, the contents of TG and LDL-C were significantly reduced, and the content of HDL-C was increased in LMH-KO-VNN1 cells. Transcriptome sequencing showed that there were 1,335 DEGs between LMH-KO-VNN1 cells and original LMH cells. Of these DEGs, 431 were upregulated, and 904 were downregulated. Gene ontology analyses of all DEGs showed that the lipid metabolism-related pathways, such as fatty acid biosynthesis and long-chain fatty acid biosynthesis, were enriched. KEGG pathway analyses showed that "lipid metabolism pathway", "energy metabolism", and "carbohydrate metabolism" were enriched. A total of 76 DEGs were involved in these pathways, of which 29 genes were upregulated (such as cytochrome P450 family 7 subfamily A member 1, ELOVL fatty acid elongase 2, and apolipoprotein A4) and 47 genes were downregulated (such as phosphoenolpyruvate carboxykinase 1) by VNN1 knockout in the LMH cells. Conclusion: These results suggest that VNN1 plays an important role in coordinating lipid metabolism in the chicken liver.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.505-513
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• 2017

Objective: This study was performed to evaluate whether ambient temperature and dietary glycerol addition affect growth performance, and blood metabolic and immunological parameters, in beef cattle. Methods: Twenty Korean cattle steers ($405.1{\pm}7.11kg$ of body weight [BW], $14.2{\pm}0.15$ months of age) were divided into a conventional control diet group (n = 10) and a 2% glycerol- added group (n = 10). Steers were fed 1.6% BW of a concentrate diet and 0.75% BW of a timothy hay diet for 8 weeks (4 weeks from July 28th to August 26th and 4 weeks from August 27th to September 26th). Blood was collected four times on July 28th, August 11th, August 27th, and September 26th. Results: The maximum indoor ambient temperature-humidity index in August (75.8) was higher (p<0.001) than that in September (70.0), and in August was within the mild heat stress (HS) category range previously reported for dairy cattle. The average daily gain (ADG; p = 0.03) and feed efficiency (p<0.001) were higher in hotter August than in September. Glycerol addition did not affect ADG and feed efficiency. Neither month nor glycerol addition affected blood concentrations of cortisol, triglyceride, or non-esterified fatty acid. Blood concentrations of cholesterol, low-density lipoprotein, high-density lipoprotein, glucose, and albumin were lower (p<0.05) on August 27th than on September 26 th, and blood phosphorus, calcium and magnesium concentrations were also lower on August 27th than on September 27th. Glycerol addition did not affect these blood parameters. Percentages of $CD4^+$ T cells and $CD8^+$ T cells were higher (p<0.05) on July 28th than on August 27th and September 26th. The blood $CD8^+$ T cell population was lower in the glycerol supplemented-group compared to the control group on July 28th and August 27th. Conclusion: Korean cattle may not be significantly affected by mild HS, considering that growth performance of cattle was better in hotter conditions, although some changes in blood metabolic and mineral parameters were observed.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2291-2295
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• 2014

Although nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis and expression has been observed in some types of human cancer and stem cells, the molecular mechanisms involved in mediation of cell proliferation and cell cycling remains largely elusive. The aim of the present study was to evaluate NS as a potential target for gene therapy of human breast carcinoma by investigating NS gene expression and its effects on SKBR-3 cell proliferation and apoptosis. NS mRNA and protein were both found to be highly expressed in all detected cancer cell lines. The apoptotic rate of the pcDNA3.1-NS-Silencer group ($12.1-15.4{\pm}3.8%$) was significantly higher than those of pcDNA3.1-NS ($7.2-12.0{\pm}1.7%$) and non-transfection groups ($4.1-6.5{\pm}1.8%$, P<0.01). MTT assays showed the knockdown of NS expression reduced the proliferation rate of SKBR-3 cells significantly. Matrigel invasion and wound healing assays indicated that the number of invading cells was significantly decreased in the pcDNA3.1-NS-siRNA group (P<0.01), but there were no significant difference between non-transfected and over-expression groups (P>0.05). Moreover, RNAi-mediated NS down-regulation induced SKBR-3 cell G1 phase arrest, inhibited cell proliferation, and promoted p53 pathway-mediated cell apoptosis in SKBR-3 cells. NS might thus be an important regulator in the G2/M check point of cell cycle, blocking SKBR-3 cell progression through the G1/S phase. On the whole, these results suggest NS might be a tumor suppressor and important therapeutic target in human cancers.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3195-3201
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• 2015

Objectives: To investigate the effects of betaine on HeLa cell growth and apoptosis and molecular mechanisms. Materials and Methods: Concentrations of 0.1, 1.0, 5.0, 20.0, 100.0 mg/ml of betaine were used to evaluate the anticancer efficacy for HeLa cells respectively, and MCF-10A was also detected as a normal diploid cell control. Results: We found that proliferation of HeLa cells was inhibited significantly upon exposure to increasing betaine levels with the MTT test (p<0.05). The percentage of S phase cells in the low dose groups (<5mg/ml) were distinctly higher than in high dose groups, and the rates of Sub-G1 phase were the opposite (p<0.01); A high concentration of betaine (>5.0mg/ml) significantly promoted the apoptosis of HeLa cells (p<0.01). SOD activities of the low dose groups were slightly higher than the control group (p<0.05) and there were obvious synchronicity and correlation among the expression of promoting apoptosis genes Bax, P53, Caspase 3 and apoptosis suppression gene Bcl-2. In response to an apoptosis-inducing stimulus, p53 and cyclin D1 could be activated with blockage of the cell cycle at G1/S or S/G2 checkpoints. Conclusions: Our data showed that betaine could promote HeLa cells proliferation in vitro at low concentrations. In contrast, high concentrations could significantly inhibit cell growth and migration, and induce apoptosis of HeLa cells through caspase 3 signaling and further promoted necrosis. This might imply that betaine exhibits tumoricidal effects and acts as a biological response modifier in cancer treatment by inducing apoptosis and cell cycle arrest in a dose and time-dependent manner.

• Journal of Ecology and Environment
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• v.33 no.4
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• pp.289-298
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• 2010

A number of characteristics of the Lake H$\ddot{o}$vsg$\ddot{o}$l watershed, such as the lake's location at the edge of the Central Asian continuous permafrost zone, provide a unique opportunity to evaluate possible anthropogenic impacts in this remote area in northern Mongolia. In this study, we compared stream solute concentrations in six sub-watersheds in the Lake H$\ddot{o}$vsg$\ddot{o}$l watershed. Water samples were collected during the summer months between 2003 and 2005. Concentrations of $Cl^-$ ranged from 9.8 to $51.3\;{\mu}mol/L$; average nitrate concentrations were very low and ranged from undetectable to $1.1\;{\mu}mol/L$ and average ${SO_4}^{2-}$ concentration at sampling stations with minimal animal grazing ranged from 66 to $294\;{\mu}mol/L$. Average dissolved organic carbon (DOC) concentrations ranged from 642 to $1,180\;{\mu}mol$ C/L. We did not find statistically significant differences in DOC concentrations among the six streams, although DOC concentrations tended to be higher in the two northernmost streams, possibly related to differences in the active layer above the permafrost. Dissolved organic nitrogen (DON) concentrations were correlated with DOC concentration, and followed the same spatial pattern as those for DOC. In streams in this remote watershed, total dissolved nitrogen was made up of mostly organic N, as has been found for other regions distant from anthropogenic N sources. Overall, these results suggest that future research on the dynamics of DOC and DON in this watershed will be especially insightful in helping to understand how changes in climate and land use patterns will affect transformations, retention, and export of dissolved organic matter within these sub-watersheds in the Lake H$\ddot{o}$vsg$\ddot{o}$l region.

• Korean Journal of Medicinal Crop Science
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• v.25 no.4
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• pp.231-237
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• 2017

Background: Cisplatin is one of the most extensively used chemotherapeutic agents for the treatment of cancer, including bladder, and ovarian cancers. However, it has been shown to induce nephrotoxicity, despite being an outstanding anti-cancer drug. In this study, we investigated the protective effect of dopaol ${\beta}$-D-glucoside (dopaol) on cisplatin-induced nephrotoxicity. Methods and Results: To confirm the protective effect of dopaol on cisplatin-induced nephrotoxicity, HK-2 cells were treated with $20{\mu}M$ cisplatin and $80{\mu}M$ dopaol. Cisplatin increased apoptosis, caspase-3 activity and mitochondrial dysfunction; however pretreatment with $80{\mu}M$ dopaol successfully attenuated apoptosis, caspase-3 activity and mitochondrial dysfunction. To evaluate the protective effect dopaol on cisplatin-induced nephrotoxicity in vivo, we used an animal model (balb/c mice, 20 mg/kg, i.p. once/day for 3 day). The results were similar to those obtained using HK-2 cells; renal tubular damage and neutrophilia induced by cisplatin reduced following dopaol injection (10 mg/kg, i.p. once/day for 3 day). Conclusions: These results indicate that dopaol treatment reduced cisplatin-induced nephrotoxicity in vitro and in vivo, and can be used to treat cisplatin-induced nephrotoxicity. However, further studies are required to determine the toxicity high dose dopaol and the signal pathways involved in its mechanism of action in animal models.

• Animal Bioscience
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• v.34 no.8
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• pp.1415-1424
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• 2021

Objective: Portable laser methane detectors (LMDs) may be an economical means of estimating CH₄ emissions from ruminants. We validated an LMD-based approach and then used that approach to evaluate CH₄ emissions from indigenous dairy cows in a dryland area of Ethiopia. Methods: First, we validated our LMD-based approach in Simmental crossbred beef cattle (n = 2) housed in respiration chambers and fed either a high- or low-concentrate diet. From the results of the validation, we constructed an estimation equation to determine CH₄ emissions from LMD CH₄ concentrations. Next, we used our validated LMD approach to examine CH₄ emissions in Fogera dairy cows grazed for 8 h/d (GG, n = 4), fed indoors on natural-grassland hay (CG1, n = 4), or fed indoors on Napier-grass (Pennisetum purpureum) hay (CG2, n = 4). All the cows were supplemented with concentrate feed. Results: The exhaled CH₄ concentrations measured by LMD were linearly correlated with the CH₄ emissions determined by infrared-absorption-based gas analyzer (r² = 0.55). The estimation equation used to determine CH₄ emissions (y, mg/min) from LMD CH₄ concentrations (x, ppm m) was y = 0.4259x+38.61. Daily CH₄ emissions of Fogera cows estimated by using the equation did not differ among the three groups; however, a numerically greater milk yield was obtained from the CG2 cows than from the GG cows, suggesting that Napier-grass hay might be better than natural-grassland hay for indoor feeding. The CG1 cows had higher CH₄ emissions per feed intake than the other groups, without significant increases in milk yield and body-weight gain, suggesting that natural-grassland hay cannot be recommended for indoor-fed cows. Conclusion: These findings demonstrate the potential of using LMDs to valuate feeding regimens rapidly and economically for dairy cows in areas under financial constraint, while taking CH₄ emissions into consideration.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.2
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• pp.151-157
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• 2005

Sheep production is affected by genetic and non-genetic factors. A knowledge of these factors is essential for efficient management and for the accurate estimation of breeding values. The objective of this study was to establish the non-genetic factors which affect birth weight and weaning weight in Dorper, Mutton Merino and indigenous Sabi sheep breeds. A total of 2,625 birth and weaning weight records from Grasslands Research Station collected from 1991 through 1993, were used. The records were collected from indigenous Sabi (939), Dorper (807) and Mutton Merino (898) sheep. A mixed classification model containing the fixed effects of year, birth status and sex was used for identification of non-genetic factors. Sire within breed was included as a random effect. Two factor interactions and three factor interactions were important in indigenous Sabi, Mutton Merino and Dorper sheep. The mean birth weights were 4.37${\pm}$0.04 kg, 4.62${\pm}$0.04 kg and 3.29${\pm}$0.04 kg for Mutton Merino, Dorper and Sabi sheep, respectively. Sire had significant effects (p<0.05) on birth weight in Mutton Merino and indigenous Sabi sheep. Year of lambing had significant effects (p<0.05) on birth weight in indigenous Sabi, Mutton Merino and Dorper sheep. The effect of birth status was non significant in Dorper and Mutton Merino sheep while effect of birth status was significant on birth weight in indigenous Sabi sheep. In Indigenous Sabi sheep lambs born as singles (3.30${\pm}$0.05 kg) were 0.23 kg heavier than twins (3.07${\pm}$0.05 kg), in Mutton Merino lambs born as singles (3.99${\pm}$0.08 kg) were 0.07 kg heavier than twins (3.92${\pm}$0.08 kg) and in Dorper lambs born as singles (4.41${\pm}$0.04 kg) were 0.02 kg heavier than twins (4.39${\pm}$0.04 kg). On average males were heavier than females (p<0.05) weighing (3.32${\pm}$0.04 kg vs. 3.05${\pm}$0.07 kg) in indigenous Sabi, 4.73${\pm}$0.03 kg vs. 4.08${\pm}$0.05 in Dorper and 4.26${\pm}$0.07 kg vs. 3.66${\pm}$0.09 kg in Mutton Merino sheep. Two way factor interactions of sire*year, year*sex and sex*birth status had significant effects (p<0.05) on birth weight in indigenous Sabi, Mutton Merino and Dorper sheep while the effect of year*birth status was non significant on birth weight in Indigenous Sabi sheep. The three way factor interaction of year*sex*birth status had a significant effect (p<0.01) on birth weight in indigenous Sabi and Mutton Merino. Tupping weight fitted as a covariate had significant effects (p<0.001) on birth weight in indigenous Sabi, Mutton Merino and Dorper sheep. The mean weaning weights were 17.94${\pm}$0.31 kg, 18.19${\pm}$0.28 kg and 14.39${\pm}$0.28 kg for Mutton Merino, Dorper and Indigenous Sabi sheep, respectively. Effects of sire and sire*year were non significant on weaning weight in Dorper and Mutton Merino while year, sex and sex*year interaction had significant effects (p<0.001) on weaning weight. On average males were heavier than females (p<0.001) at weaning. The respective weaning weights were 18.05${\pm}$0.46 kg, 18.68${\pm}$0.19 kg, 14.14${\pm}$0.15 kg for males and 16.64${\pm}$0.60 kg, 16.41${\pm}$0.31 kg, 12.64${\pm}$0.32 kg for females in Mutton Merino, Dorper and Indigenous Sabi sheep. Lambs born as singles were significantly heavier at weaning than twins, 0.05 kg, 0.06 kg and 0.78 kg for Mutton Merino, Dorper and Indigenous Sabi sheep, respectively. Effect of tupping weight was highly significant on weaning weight. The three way factor interaction year*sex*birth status had a significant effect (p<0.01) on weaning weight. Correction for environmental effects is necessary to increase accuracy of direct selection for birth weight and weaning weight.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.2
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• pp.161-168
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• 2014

This study aimed to investigate dietary concentrate:forage ratios (C:F) and undegraded dietary protein (UDP) on nitrogen balance and urinary excretion of purine derivatives (PD) in lambs. Four Dorper${\times}$thin-tailed Han crossbred castrated lambs with $62.3{\pm}1.9$ kg body weight at 10 months of age were randomly assigned to four dietary treatments in a $2{\times}2$ factorial arrangement of two levels of C:F (40:60 and 60:40) and two levels of UDP (35% and 50% of CP), according to a complete $4{\times}4$ Latin-square design. Each experimental period lasted for 19 d. After a 7-d adaptation period, lambs were moved into individual metabolism crates for 12 d including 7 d of adaption and 5 d of metabolism trial. During the metabolism trial, total urine was collected for 24 h and spot urine samples were also collected at different times. Urinary PD was measured using a colorimetric method and creatinine was measured using an automated analyzer. Intake of dry matter (DM) (p<0.01) and organic matter (OM) (p<0.01) increased as the level of UDP decreased. Fecal N was not affected by dietary treatment (p>0.05) while urinary N increased as the level of UDP decreased (p<0.05), but decreased as dietary C:F increased (p<0.05). Nitrogen retention increased as dietary C:F increased (p<0.05). As dietary C:F increased, urinary excretion of PD increased (p<0.05), but was not affected by dietary UDP (p>0.05) or interaction between dietary treatments (p>0.05). Daily excretion of creatinine was not affected by dietary treatments (p<0.05), with an average value of $0.334{\times}0.005$ mmol/kg $BW^{0.75}$. A linear correlation was found between total PD excretion and PDC index ($R^2$ = 0.93). Concentrations of creatinine and PDC index in spot urine were unaffected by sampling time (p>0.05) and a good correlation was found between the PDC index (average value of three times) of spot urine and daily excretion of PD ($R^2$ = 0.88). These results suggest that for animals fed ad libitum, the PDC index in spot urine is effective to predict daily excretion of PD. In order to improve the accuracy of the spot sampling technique, an appropriate lag phase between the time of feeding and sampling should be determined so that the sampling time can coincide with the peak concentration of PD in the urine.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.6
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• pp.766-773
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• 2011

The objective of the present study was to examine whether yaks possess any adaptive mechanisms of nitrogen (N) metabolism to survive in the harsh foraging environment of the Qinghai-Tibetan Plateau. A grazing experiment on native alpine meadows was conducted to determine availability of herbage biomass and body weight (BW) change of yaks over the year, followed by two indoor feeding trials to investigate adaptation mechanisms of N metabolism in yaks fed at similar intake level to grazing conditions. Three castrated males of each of three genotypes; yak (Bos grunniens), indigenous cattle (Bos taurus) and their crossbred - cattleyak (Bos taurus male${\times}$Bos grunniens female), were used in the housed trials. Results showed that: i) Monthly herbage biomass production and daily grazing intakes by yaks over the year ranged from 220 to 4,664 kg DM per ha, and 1.90 to 8.50 kg DM, respectively. For about seven months each year, yaks suffer from malnutrition as a result of inadequate pasture conditions; ii) Urinary N excretion and N retention by yaks were significantly affected by feeding level, and yaks had a lower (p<0.05) average daily urinary N excretion (0.39 g/kg $BW^{0.75}$) and a greater (p<0.05) N retention (-0.09 g/kg $BW^{0.75}$) than indigenous cattle (0.47 and -0.16 g/kg $BW^{0.75}$, respectively). Fasting daily urinary N excretion was greater (p<0.05) for indigenous cattle than yaks (353 vs. 248 mg/kg $BW^{0.75}$). Purine derivative N excretion and purine derivative N index (PNI) increased with increasing feeding level, while the value of PNI was greater (p<0.05) for yaks and cattleyak (0.11 and 0.12, respectively) than for indigenous cattle (0.09) during the feeding trials. These results suggest that yaks could rely, in part, on the recycling of N to adapt to the harsh forage environment on the Qinghai-Tibetan Plateau.), were used in the housed trials. Results showed that: i) Monthly herbage biomass production and daily grazing intakes by yaks over the year ranged from 220 to 4,664 kg DM per ha, and 1.90 to 8.50 kg DM, respectively. For about seven months each year, yaks suffer from malnutrition as a result of inadequate pasture conditions; ii) Urinary N excretion and N retention by yaks were significantly affected by feeding level, and yaks had a lower (p<0.05) average daily urinary N excretion (0.39 g/kg $BW^{0.75}$) and a greater (p<0.05) N retention (-0.09 g/kg $BW^{0.75}$) than indigenous cattle (0.47 and -0.16 g/kg $BW^{0.75}$, respectively). Fasting daily urinary N excretion was greater (p<0.05) for indigenous cattle than yaks (353 vs. 248 mg/kg $BW^{0.75}$). Purine derivative N excretion and purine derivative N index (PNI) increased with increasing feeding level, while the value of PNI was greater (p<0.05) for yaks and cattleyak (0.11 and 0.12, respectively) than for indigenous cattle (0.09) during the feeding trials. These results suggest that yaks could rely, in part, on the recycling of N to adapt to the harsh forage environment on the Qinghai-Tibetan Plateau.