• Clinical and Experimental Reproductive Medicine
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• v.36 no.1
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• pp.23-34
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• 2009

Objectives: Mesenchymal stem cells (MSC) comprise a promising tool for cellular therapy. It is known that long-term in vitro culture of human bone marrow and adipose tissue derived-MSCs lead to a reduction of life span and a change of stem-like characters. The aim of our study was to examine whether stem cell properties of human umbilical cord-derived stem cells (HUC) could be affected by in vitro expansion. Methods: HUC were isolated from human umbilical cord and cultured for 10 passages in vitro. Morphology and population doubling time (PDT) were investigated, and changes of stem cell properties were examined using RT-PCR and immunocytochemistry during serial subcultures. Results: Morphology and PDT of HUC began to change slightly from the 7th passage (p7). Expression level of nestin and vimentin mRNAs increased along with the culture period from p4 until p10. In contrast, expression level of SCF mRNA decreased during the same culture period. Expression level of Oct-4 and HNF-4${\alpha}$ mRNAs was not significantly changed throughout the culture period until p10. Expression level of BMP-4, FGF-5, NCAM and HLA-ABC mRNAs appeared to increase as the culture continued, however, the difference was not significant. Immunocytochemical studies showed that HUC at p3, p6 and p9 positively were stained with antibodies against SSEA-3 and SSEA-4 proteins. Interestingly, staining intensity of HUC for ICAM-1 and HLA-ABC gradually increased throughout the culture period. Intensity against thy-1 and fibronectin antibodies increased at p9 while that against TRA-1-60 and VCAM-1 antibodies began to decrease at p6 until p9. Conclusions: These results suggest that HUC change some of their stem cell characteristics during in vitro culture. Development of culture system might be needed for the maintenance of characteristics.

• Applied Microscopy
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• v.37 no.4
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• pp.271-280
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• 2007

Tonicity-responsive enhancer binding protein(TonEBP) is a transcriptional factor essential in the function and development of the renal medulla. TonEBP plays a critical role in protecting renal medullary cells from the deleterious effect of hypertonicity. TonEBP is a key regulator of urinary concentration via stimulation of transcription of urea transporter(UT) in a manner independent of vasopressin. UT in the renal inner medulla is important for the conservation of body water due to its role in the urine concentrating mechanism. Mongolian gerbil(Meriones unguiculatus) has been as an model animal for studying the neurological disease such as stroke and epilepsy because of the congenital incomplete in Willis circle, as well as the investigation of water metabolism because of the long time-survival in the condition of water-deprived desert condition, compared with other species animal. In this study, we divide 3 groups of which each group include the 5 animals. In the study of 7 or 14 days water restricted condition, we investigated the TonEBP and UT-A by using a immunohistochemistry in the kidney. In the normal kidney, the distribution of TonEBP is generally localized on nuclei of inner medullary cells. Nuclear distribution of TonEBP is generally increased throughout the medulla in 7 and 14 days dehydrated group compared with control group. Increased nuclear localization was particularly dramatic in thin limbs. In control groups, UT-A was expressed in inner stripe of outer medulla(ISOM) and inner medulla(IM). UT-A was present in the terminal part of the short-loop of descending thin limbs (DTL) in ISOM and also present in the inner medullary collecting duct(IMCD), where the intensity of it gradually increased toward the papillary tip. In the dehydrated kidney, UT-A immunoreactivity was increased in the short-loop of DTL in ISOM and in the long-loop of DTL in the initial part of IM, where was expressed moderate positive reaction in the normal kidney. Also it was up regulated in the IMCD in initial & middle part of IM. However UT-A down regulated in the IMCD, where the intensity of it gradually decreased toward the papillary tip. These findings suggest that increased levels of TonEBP in medulla and UT-A in shot-loop of DTL and IMCD play a important role for maintain fluid balance in the water-deprived mongolian gerbil kidney.

• Korean Journal of Poultry Science
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• v.30 no.3
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• pp.197-202
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• 2003

Feeding probiotics in broiler chicks still critical in several aspects. Thus, this study was conducted to investigate the impact of feeding multiple probiotics on performance, intestinal microflora, blood cholesterol and ND antibody vaccine titer in broiler chicks. Three hundred twenty one day old male broiler chicks(cobb ${\times}$ cobb) were divided into four levels of multiple probiotics(0, 0.1, 0.2, 0.3%) with five replicates for 35 days. Basal diets contained 21.5, 19.0% CP and 3,100 kcal/kg ME for starting and finishing period, respectively. Weight gain, feed intake and feed conversion were measured weekly. The number of Salmonella, E. coli, Lactobacillus, and yeast were examined from ileum and cecum at the end of experiment. ND vaccine titer, cholesterol were detected from sera. Weight gain of birds fed probiotics were 669.33, 679.75 at the level of 0.1 and 0.2% supplemental groups for starting period. It was also improved in those treatments for finishing period and higher than control for total period. Feed conversion tended to be improved compared to that of control by the supplementation of probiotics for the first three weeks and seemed to show the similar tendency for the rest of two weeks. It was 1.611, 1.621 for the entire feeding period and improved compared with control. Total salmonella, was not decreased in ileal digesta of birds fed the probiotics compared with control, whereas the number of yeast increased in 0.1% treatment. However, the number of Lactobacillus and yeast in cecum was higher than control. Even though the blood cholesterol seem to high in 0.1% probiotics treatment, the ratio of HDL to total cholesterol showed higher than control. ND vaccine titer of birds fed probiotics were significantly higher than control (P<0.05). These results 0.1% multiple probiotics would be possible to improve the performance of broiler chicks and ND vaccine titer.

• Korean Journal of Poultry Science
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• v.35 no.3
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• pp.283-290
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• 2008

The Siberian ginseng and Eucommia are a kind of medicinal plant with powerful anti-oxidant activity. An experiment was conducted to investigate the effect of Siberian ginseng leaf and Eucommia leaf at level of 0.5% and 1% per feed in Ross commercial broiler for 4 to 35 days of age on performance, organ weight, blood biochemical profiles and telomere quantity. Chickens consuming diets containing 1% Siberian ginseng had higher feed conversion ratio than the other treated chicken during experimental period whereas no significant differences were detected in body weight, weight gain and feed intake. The weight of bursa of fabricius was significantly increased in chickens with dietary supplementation compared with chickens fed control but this was not seen in liver, spleen and thymus. In blood biochemical profiles, chickens with dietary supplementation had higher concentration than chickens fed control in triglyceride, cholesterol and glucose. The concentration of aspartate aminotransferase, alanine aminotransferase, albumin and total protein, however, was not significantly different between dietary supplemented chickens and control chickens. The relative amount of telomeric DNA of lymphocytes in chickens with dietary supplementation was significantly higher than that of control chickens but the difference was not found in liver, heart and testis tissues. In conclusion, dietary supplementation of Siberian ginseng and Eucommia in broiler improved immune activity and telomere length without decreasing chicken growth performance.

• Journal of the Korean Applied Science and Technology
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• v.36 no.3
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• pp.1018-1027
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• 2019

Natural-derived protein-derived low molecular weight peptides have been known to have physiological activities such as antioxidant, hypertension relief, immunomodulation, pain relief and antimicrobial activity. In this study, the low-molecular peptides were produced using commercial proteases (alcalase, bromelain, flavourzyme, neutrase, papain, protamex), and the antioxidant activity (DPPH scavenging activity, superoxide radical scavenging activity, hydroxy radical scavenging activity, and metals chelation capacity), constituent amino acid and molecular weight of the peptide were analyzed. Enzyme reaction was performed by adding 50 g of chopped Ogae meat slurry and 2%(w/v) protein enzyme into the enzyme reactor for 2 h at a pH of 6 and a temperature of $60^{\circ}C$. The degree of hydrolysis(%) after the reaction ranged from $36.65{\pm}4.10%$ to $70.75{\pm}5.29%$. The highest degree of hydrolysis of protamex was 46.3%, and the highest value of papain hydrolysate was $70.75{\pm}5.29%$. On the other hand, alcalase hydrolysate showed the lowest value of $36.65{\pm}4.10%$. Bromelain-treated low molecular weight peptides showed the highest DPPH radical scavenging activity and the lowest scavenging activity of alcalase-treated peptides. Superoxide radical scavenging activity showed that bromelain treated low molecular peptide showed the highest radical scavenging activity of 50% or more. Hydroxyl radical scavenging activity ranged from about 16.73 to 69.16%, the highest among bromelain-treated low molecular peptides. $Fe^{2+}$ chelation abilities showed a distribution between about 17.85 to 47.84%. The chelation capacity of the hydrolysates was not significantly different without any difference to the enzymes used. The results of amino acid analysis showed differences between hydrolysates of alcalase, bromelain, flavourzyme, neutrase, papain, and protamex enzymes. The most amino acid was glutamic acid. The molecular weight distribution of the enzyme hydrolyzates was in the range of 300-2,000 Da, although the molecular weight distribution differed according to the treated enzymes.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.4 no.1
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• pp.37-53
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• 1998

The sprig of Jinryungtang Gagambang has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Jinryungtang Gagambang extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell liens (A549, hep3B, Caki-1, Sarcoma 180) after exposure to Jinryungtang Gagambang extract using in ILS, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Jinryungtang Gagambang extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Jinryungtang Gagambang extract showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Jinryungtang Gagambang decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Jinryungtang Gagambang also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Jinryungtang Gagambang extract into TBM. 4. Jinryungtang Gagambang extract also increased NK cell activities. According to the above results, it could be suggested that Jinryungtang Gagambang extract has prominent antiutmor effect.

• Korean Journal of Poultry Science
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• v.7 no.2
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• pp.54-76
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• 1980

A survey was conducted to determine the status of health and productivity of poultry farms in Korea. Area included Was Kyunggido where exist nearly 50% of national poultry population. From this area, 41 layer and 34 broiler farms covering 21 Countries were selected randomly for the survey. When farms were divided in the operation size, 95.1% of layer and 82.3% of broiler farms were classified as business or industrial level while the rest were managed in a small scale as part time job. Generally layer farms had been established much earlier than broiler farms. Geographically 10.7% of layer farms were sited near the housing area such as field foreast and rice field. No farms were located near the seashore. The distance from one farm from the other was very close, being 80% of the farms within the distance of 1km and as many as 28% of the farms within loom. This concentrated poultry farming in a certain area created serious problems for the sanitation and preventive measures, especially in case of outbreak of infectious diseases. Average farm size was 5,016${\times}$3.3㎡ for layers and 1,037${\times}$3.3㎡ for broilers. 89.5% of layer ana 70.6% of broiler farms owned the land for farming while the rest were on lease. In 60% of layer farms welters were employed for farming while in the rest their own labour was used. Majority of farms were equipped poorly for taking necessary practice of hygiene and sanitation. The amount of disinfectant used by farms was considerably low. As many as 97.6% of lave. farms were practised with Newcastle(ND) and fowl pox(F$.$pox) vaccine, whereas only 43.6% and 5.1% of broiler farms were practised with ND and F$.$pox vaccine, respectively. In 17-32.7% of farms ND vaccine was used less than twice until 60 days of age and in only 14.6% of farms adult birds were vaccinated every 4months. Monthly expense for preventive measures was over 200,000W in 32% of farms. Only 4.9-2.7% of vaccine users were soaking advice from veterinarians before practising vaccination, 85% of the users trusted the efficacy of the vaccines. Selection of medicine was generally determined by the farm owner rather than by veterinarans on whom 33.3% of farms were dependant. When diseases outbroke, 49.3% of farms called for veterinary hospital and the rest were handled by their own veterinarians, salesmen or professionals. Approximately 70% of farms were satisfied with the diagnosis made by the veterinarians. Frequency of disease outbreaks varied according to the age and type of birds. The livabilities of layers during the period of brooding, rearing ana adultwere 90.5, 98.9 and 75.2%, respectively while the livalibility of broilers until marketing was 92.2%. In layers, average culling age, was 533.3 day and hen housed eggs were 232.7. Average feed conversion rates of layers and broilers were 3.30 and 2.48, respectively. Those figures were considerably higher than anticipated but still far lower than those in developed countries.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.937-946
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• 2005

This experiment was conducted to investigate the effects of dietary Bacillus subtilis on growth performance, nutrient digestibility, immunological cells change, fecal noxious gas and carcass meat quality characteristics in finishing pigs. The dietary treatments were 1) CON(basal diet), 2) BS0.1(basal diet+ 0.1% Bacillus subtilis) and 3)BS0.2(basal diet+0.2% Bacillus subtilis). Sixty crossbred(Landrace$\time$ Yorkshire$\time$Duroc) pigs(89.5$\pm$0.11kg average initial body weight) were used in a 42 days growth trial. The pigs were assigned to the treatments according to body weight and each treatment had 5 replicates of 4 pigs per pen in a randomized complete block design. Through the entire experimental period, average daily gain(ADG) and average daily feed intake(ADFI) were not significantly different among the treatments. Pigs fed BS0.1 diet significantly increased their gain/feed compared to pigs fed CON and BS0.2 diets(P<0.05). Also, dry matter(DM) and nitrogen(N) digestibilities were greater in the pigs fed BS0.1 diet than those fed CON diet(P<0.05). There were no significant differences in fecal NH3-N concentration among the treatments. In blood assay for immunological cells change investigations, red blood cells(RBC) counts increased in the pigs fed BS0.2 diet compared to pigs fed CON and BS0.1 diets. There were no significant differences in carcass pH, drip loss, marbling and firmness. However, sensual color and a*(redness) value of meat in the pigs fed BS0.2 diet were higher than in pigs fed CON diet(P<0.05). Therefore, this experiment suggested that Bacillus subtilis supplementation could improve nutrient digestibility, RBC counts and carcass meat color of pigs.

• IMMUNE NETWORK
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• v.1 no.1
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• pp.87-94
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• 2001

Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.

• Journal of Life Science
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• v.18 no.12
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• pp.1693-1699
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• 2008

Recent studies have suggested that inulin might be utilized as a prebiotics for the promotion of antimicrobial growth, but a major obstacle to the use of inulin has been its low bifidogenic effects, which were initially observed in the ceca of broiler chickens. Inulin has some problems with related to denaturation in air and lowering passage rate from upper digestive tract to caecum. To solve this problems, a newly developed compound derived by microencapsulation, inuloprebiotics, was hypothesized to enrich cecal bifidobacterial populations and reduce the colonization levels of Salmonella in the ceca of broiler chickens. The in vitro growth of intestinal beneficial bacteria including Bifidobacterium longum, Bifidobacterium bifidum, Lactobacillus acidophilus, and Lactobacillus casei grew effectively on the medium containing inulin, whereas the growth of Streptococcus aureus and Clostridium perfringens was not differences among the treatment groups. Broiler chickens consumed chow diets containing 0.5%, 0.7% or 1.0% inuloprebiotics, or a control diet without inuloprebiotics supplementation. The chickens on the inuloprebioticssupplemented diets evidenced significantly higher cecal levels of Bifidobacterium and Lactobacillus species as compared with the chickens on the control diet. The population of cecal E. coli and Salmonella was specifically reduced as the result of treatment with inuloprebiotics. However, we noted no significant differences in Bifidobacterium species, E. coli and Salmonella counts among the inuloprebiotics treatment groups. The inuloprebiotics-supplemented diets induced an increase in the serum IgG concentration. The thymus index was significantly increased in the broiler chickens that consumed diets containing 0.7% or 1.0% inuloprebiotics, with the exception of the chickens consuming the diet supplemented with 0.5% inuloprebiotics. These results indicate that the inuloprebiotic preparations exerted an immune system-promoting effect or selectively enriched the cecal Bifidobacterium species populations in the broiler chickens, and also suggest that inuloprebiotics may prove useful as a stable natural antimicrobial agent.