• Journal of Life Science
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• v.18 no.3
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• pp.344-349
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• 2008

Normal somatic cells proliferate for a limited number of doublings in culture and then enter an irreversible growth-arrest state called replicative senescence. Replicative senescence has been believed a reason for the limited cellular turnover and deterioration of tissue function in aged animals. However, there is no experimental evidence supporting this assumption. Furthermore, cells from aged person have been poorly characterized with an exception of the cases of T cells. In this study, we examined cell biological changes occurring in replicative senescence of fibroblast strains originated from a new-born (NHF-NB) and a 87 year old man (NHF-87). NHF-87 (and the cells from a 75-year old) proliferated to smaller population doublings and with longer doubling times than NHF-NB did. At early passages, NHF-87 exhibited a low senescence-associated ${\beta}-Gal$ (SA ${\beta}-Gal$) activity and lipofuscin level, typical markers for cellular senescence. Furthermore, they maintained low levels of lysosome and reactive oxygen species (ROS). All of these levels increased dramatically in the late passage NHF-87 quite similarly as those in the late passaged NHF-NB did. These results indicate that most cells originated from the aged maintain a phenotype of the cells originated from new-born donors and undergo replicative senescence with the same kinetics as that of the cells from new-born. It is also indicated that not SA ${\beta}-gal$ activity but cell proliferation rate may be qualified as a biomarker for cells aged in vivo.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.207-212
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• 2006

The aim of this study was to examine the effect of mechanical enucleation methods, aspiration and squeezing, on the developmental ability of nuclear transfer bovine embryos. Enucleated oocytes made by both enucleation methods were fused to adult ear skin cells. After 7 days of culture, developmental ability up to blastocyst stage was similar in both squeezing($33.6{\pm}15.7%$) and aspiration enucleation methods($31.9{\pm}13.4%$). The proportion of blastocysts at Day 8 of culture was also similar between the aspiration($37.8{\pm}10.4%$) and squeezing enucleatign s($35.3{\pm}15.1%$). The mean cell number in Day 7 blastocysts was also similar between the both groups(aspiration: $110.3{\pm}39.2$ vs. squeezing: $103.7{\pm}42.8$). The ratio of apoptotic cells was also found to be not significant different between the both groups(aspiration: $2.8{\pm}2.6%$ vs. squeezing: $4.3{\pm}4.4%$). These results suggest that aspiration and squeezing methods, as mechanical enucleation technique, are both useful for the production of bovine somatic cell nuclear transfer embryos.

• Journal of Animal Science and Technology
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• v.50 no.5
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• pp.705-712
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• 2008

Sixteen multiparous lactating Holstein cows were used to compare effects of supplementing 1)no additive(Control), 2)1.2% sodium bicarbonate(NaHCO3); 3)niacin(80g/d), 4)vitamin A+E (140,000IU+1000IU) on feed intake, milk production, milk composition and somatic cell counts during the summer months. Insofar as possible, treatment groups were balanced for lactation number and days in milk. Cows were fed a diet of 9.1kg DM of concentrate and 10.2kg DM of corn silage. Daily maximum air temperature in free stall barn was 35℃ for 3 days of the pretreatment periods and decreased gradually up to 27℃ during the treatment periods of 15days. Dry matter intake of corn silage was higher(p<0.05) for cows consuming NaHCO3 than those not consuming NaHCO3. Daily milk production for niacin and vitamin A+E supplementations resulted in significant(p<0.001) increase in milk production from 3 day of trials than control and NaHCO3. Milk fat percentage tended(p=0.09) to increase and milk lactose percentage was increased significantly(p<0.001) for cows supplemented with NaHCO3, niacin and vitamin A+E. Milk protein percentages was higher significantly(p<0.05) with supplemental niacin and somatic cell counts was higher significantly(p<0.001) with supplemental vitamin A+E. These data strongly suggest that supplementation of NaHCO3, niacin or vitamin A+E should be increased for improving milk production and mammary gland health of dairy cows under heat stress.

• Applied Microscopy
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• v.40 no.2
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• pp.89-99
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• 2010

Fibroblasts are the most common cells in connective tissue and are responsible for the synthesis of extracellular matrix components. The fibrosis associated with chronic inflammation and injury may contribute to cholangiocarcinoma pathogenesis, particularly through an increase in extracellular matrix components, which participate in the regulation of bile duct differentiation during development. Mitochondria produce ATP through oxidative metabolism to provide energy to the cell under physiological conditions. Also, mitochondrial dysfunction and oxidative stress have been implicated in cellular senescence and aging. Alternations in mitochondrial structure and function are early events of programmed cell death or apoptosis and mitochondria appear to be a central regulator of apoptosis in most somatic cell. Clonorchis sinensis, one of the most important parasite of the human bile duct in East Asia, arouses epithelial hyperplasia and ductal fibrosis. Isolated fibroblast from the bile ducts of rats infected by C. sinensis showed increase of cytoplasmic process. In addition, decrease of cellular proliferation was observed in fibroblasts which was isolated from normal rat bile duct and then cultured in media containing C. sinensis excretory-secretory product. However, the effects of C. sinensis infection on the mitochondrial enzyme distribution is not clearly reported yet. Therefore, we investigated the structural change of C. sinensis infected bile duct and mitochondrial enzyme distribution of the cultured fibroblast isolated from the C. sinensis infected rat bile duct. As a result, C. sinensis infected SD rat bile ducts showed the features of chronic clonorchiasis, such as ductal connective and epithelial tissue dilatation, or ductal fibrosis. In addition, fibroblast in ductal connective tissue was damaged by physical effect of fibrotic tissue and chemical stimulation. Immunohistochemically detected mitochondrial electron transferase (ATPase, COXII, Porin) was decreased in C. sinensis infected rat bile duct and cultured fibroblast from infected rat bile duct. It can be hypothesized that the reason why number of electron transferase decrease in fibroblast isolated from the rat bile duct infected with C. sinensis is because dysfunction of electron transport system is occurred mitochondrial dysfunction, increase of ROS (reactive oxygen species) and apoptosis after chemical damage on the cell caused by C. sinensis infection. Overall, C. sinensis infection induces fibrotic change of ductal connective tissue, mutation of cellular metabolism in fibroblast and mitochondrial dysfunction. Consequently, ductal fibrosis inhibits fibroblast proliferation and decreases mitochondrial electron transferase on fibroblast cytoplasm. It was assumed that the structure of bile duct could not normalized and ductal fibrosis was maintained for a long period of time according to fibroblast metamorphosis and death induced by mitochondrial dysfunction.

• Journal of agriculture & life science
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• v.45 no.6
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• pp.135-140
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• 2011

This study was conducted to determine the effect of grape by-product on milk yield, milk composition and blood metabolites of lactating cows. A total of 8 Holstein dairy cows (average age = 89.9 months, average lactation day = 164.4, Reproduction number = 2.8) were randomly allocated into control (no additives, CON) or grape by-product treatment (add 1 kg of grape by-product per day, GBP) for 3 weeks. The concentrations of fat, protein, lactose and total solid in milk were not affected by grape by-product supplementation. However, milk yield (23.13 vs. 21.16 kg/d) was higher (p<0.05) in GBP than in CON, while SCC was lower (p<0.05). The concentrations of glucose, alanine aminotransferase, total protein and BUN in blood did not differ between treatments. But blood aspartate aminotransferase and total cholesterol concentrations (123.75 vs. 95.34 IU/L, 200.82 vs. 180.63 mg/dl) were higher (p<0.05) in cows fed GBP than CON. As the result, grape by-product, which has a various potential benefits, could be used as a functional diet supplement for lactating dairy cows.

• Journal of Aquaculture
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• v.20 no.1
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• pp.47-50
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• 2007

The primordial germ cells of the oblong rockfish, Sebastes oblongus were buried under fibrous mesenchymal tissues between gut and mesonephric duct of pre-larva with 4.0 mm total length (TL) at 1 day after the parturition. In the juvenile of 22.0 mm TL at 71 days after the parturition, the gonad composed of a large number of gonial cell and formed of cavity along the lateral side of the gonad, differentiated to the ovary. At this time, the gonad formed seminiferous tubules by somatic cells, was differentiated to the testis. The smallest oblong rockfish that possessed primary oocytes was about 42.1 mm TL at 141 days. Spermatogonia remained quiescent until most fish were over 42.1 mm TL at 141 days. The oblong rockfish the differentiated directly into male or female without an intermediate female phase at the early indifferentiated stage. Therefore, the oblong rockfish belongs to the differentiated type of gonochoristic teleosts.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.4
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• pp.305-312
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• 1992

This experiment was carried out to investigate seed set, variation in chromosome number, and agronomic characteristics of the progeny in the cross between hexaploid triticale variety, Sinkihomil(P$_1$) and tetraploid rye variety, Dooroohomil(P$_2$). Seed set rate obtained was 30.5% in the cross of Sinkihomil with Dooroohomil, whereas 3.26% in reciprocal cross using Dooroohomil as female. Alsoseed set was 8.75% in F$_1$/P$_1$, 7.20% in F$_1$/P$_2$, and 1.53% in F$_2$(=F$_1$ /F$_1$, respectively. Germination rate of crossed seed was 37% in cross of P$_1$ with P$_2$, 39.0% in F$_1$/P$_1$(BC$_1$), 50% in F$_1$/P$_2$(BC$_2$) and 43.0% in F$_1$/F$_1$(F$_2$), and 1,000 grain wight was 20.7g in the cross of P$_1$ with P$_2$, which have 41.9g and 47.7g, respectively, 24.5g in F$_1$/P$_1$, 23.6g in F$_1$/P$_2$, and 24.5g in F$_1$/F$_1$, respectively. In pollen fertility of F$_1$ plant, 69.8% turned out to be abnormal or sterile pollen grains, whereas 30.2% was fertile or normal. In meiosis of pollen mother cell of F$_1$ plant, 13.5 univalents, 8.89 bivalent and 1.24 trivalent were appeared. Somatic chromosome number of 35 in F$_1$, both 32 to 33 and 35 to 36 in F$_2$, 35 to 39 in BC$_1$ and 28 to 36 in BC$_2$ which mean producing female gamate was 14 to 18 chromosome in PMC of F$_1$ plant. Rate of fertile plant turned out to be 100% in F$_1$, 4.5% in F$_2$, 42.9% in BC$_1$, and 50.0% in BC$_2$, respectively. Number of seed set per spike appeared to be 1.17 in F$_1$ plant, 13.3 in F$_2$, 2.36 in BC$_1$, and 3.75 in BC$_2$, respectively. Days to heading of F$_1$ was intermediate, but F$_2$ was later than both parents. Plant height of F$_1$ , BC$_1$ ,and BC$_2$ was shorter than both parent, but F$_2$, longer than both parents.

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.53-58
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• 2006

The study was conducted to evaluate the effects of culture period and fusion method on the development of somatic cell nuclear transfer (SCNT) embryos reconstituted with Korean bovine fetal fibroblast cells (KbFF) and Korean bovine adult ear skin fibroblast cells (KbESF). KbFF were isolated from a day 51 Korean cattle (Hanwoo) fetus, and KbESF were isolated from a 28 month old Hanwoo calf. The cells were cultured up to 15 weeks (passage 15) in vitro for SCNT. Chamber and electrode needles were used for comparing fusion of reconstituted eggs. The doubling times of KbFF and KbESF were 17.3 hr and 24.3 hr, respectively. The fusion and cleavage rates were significantly higher in needle group (76.1 and 81.2% respectively, P<0.05) than those in chamber group. However, the blastocyst development rate was not different between both groups. Fusion and cleavage rates of NT eggs reconstituted with KbESF did not affected by passage number, however, blastocyst rates were lower in passage $1{\sim}4$ group (21.3%) than passage $5{\sim}8$ (39.4%) and $13{\sim}15$ groups (40.4%, P<0.05). Whereas, fusion rate was lower in passage $1{\sim}4$ group (61.5%) than those of passage $5{\sim}8$(75.0%) and $13{\sim}15$ (76.8%) groups, but cleavage and blastocyst rates were similar regardless of passage number in the KbFF. The results suggest that fusion method can affect the development of SCNT embryos, whereas the long term culture up to 15 passages may not affect the development of SCNT embryos.

• Korean Journal of Organic Agriculture
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• v.21 no.3
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• pp.391-401
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• 2013

This study was conducted to investigate feeding management and milk production in domestic organic dairy farms and find out ways to overcome difficulties in organic dairy farms. The number of domestic organic dairy farms was 35, which representing 0.57% among 6,068 of total dairy farms in 2011. Eleven farms among 35 organic dairy farms were surveyed. Average total raising head was 142, composed of 69 milk cow, 13 dry cow and 60 heifer and calf. The ratio of cow replacement was 42.4% in surveyed organic dairy farms. Among surveyed farms, 14.3% showed under 20kg of milk production, 57.1% represented 25~30kg of milk and 28.5% produced more than 30kg of milk. Average milk fat percentage in surveyed organic farms was 3.3%, which was lower than 4.04% milk fat percentage of whole country (2010). Based on bacterial counts (5,775 CFU/ml) and somatic cell counts (192,500 number/ml), milk quality appeared excellent in surveyed farms. 90% of surveyed farms agreed that organic milk production increased income. Among reasons for switching to organic dairy farm, environment-friendly farm management was the highest reason (54.5%). However, 45.5% of surveyed farms suffered difficulty in supply of organic feed. Therefore, stable supply of organic feed will be necessary to expand organic dairy farm in the future.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.139-143
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• 2007

This study was performed to investigate the reproductive characteristics of the cloned Hanwoo bulls produced by SCNT. The semen ejaculated from the cloned bulls (C-38 and C-39) and normal Hanwoo bull was properly measured the volume, the number of sperm, and the viability of frozen-thawed sperm. The sperm activity was analyzed using computer assisted sperm analysis (CASA). To analyze fertilizing ability of the cloned bulls, in vitro fertilization and artificial insemination were performed using the frozen-thawed semen. There were no differences in semen volume, sperm concentration, and the viability of frozen-thawed sperm between cloned bulls and normal bull. The difference was statistically significant in total motility, curvilinear velocity (VCL), straight-line velocity (VSL), and average-path velocity (VAP) of both cloned bulls compared to those of normal Hanwoo bull, respectively (p<0.05). The cleavage and blastocyst development rate were not different between the groups. five cloned cows were artificially inseminated using the frozen-thawed semen of C-38, two of them became pregnant. Two second generation calves (one male and one female) were produced. Based on these results, the cloned Hanwoo bulls showed normal reproductive abilities of semen parameters and sperm activity to their comparators and produced cloned calves, although there are some individual differences on the parameters.