In this study, anti-cancer activities of peel of Citrus junos (CJP) and Poncirus trifoliata (PTP) on MCF-7 breast cancer cells, and anti-proliferative effects of cancer cells induced by environmental hormones were investigated. The ethanol extracts of CJP and PTP inhibited cancer cell growth and induced apoptosis at the concentration over 300 mg/ml treatment for 72 hr. Morphological change of MCF-7 breast cancer cells were observed treated with the CJP and PTP of 500 mg/ml concentration for 72 hr, and apoptosis was induced by activation of caspase-3. The proliferation of MCF-7 breast cancer cells was decreased in a dose-dependent manner treated with various concentration of CJP and PTP, when compared with the control at 300 mg/ml, the proliferation of the MCF-7 breast cancer cells of both extracts was decreased over 70% and 80%, respectively.
Porcine respiratory disease complex (PRDC) is a common respiratory disease in nursery and grow-finishing pigs. A complex of viral and bacterial agents is known to be involved in the etiology of PRDC. The purpose of this study was to investigate common etiologic agents associated with PRDC in the field and compare detection methods for identifying these agents. To understand the mechanism of polymicrobial infection in PRDC, changes in the expression of cytokines were investigated. In 461 pig samples examined, most of the affected pigs ranged from 3 to 10 weeks old (73.4%), and 348 (75.4%) samples were confirmed as polymicrobial infection. Of the polymicrobial-infected cases, two (50.3%), three (32.2%), four (13.8%), five (3.2%), and six (0.5%) agents were detected. Two- or three-agent infections were the most common, with PRRSV/PCV-2 (44.6%) the most common two-agent infection. PRRSV/PCV-2/H. parasuis (11.0%) was the most common three-agent infection. Comparison of two detection methods (PCR and IHC) in the polymicrobial cases showed that 78.4% were PCV-2 positive with the PCR method, and 26.2% were PCV-2 positive with IHC. SIV was 7.8% by the PCR method and 3.7% positive by the IHC. This result indicates that the PCR method is more useful than IHC for detecting causative agents in PRDC. In the analysis of cytokines in the two- and three-agent infected samples, interleukin (IL)-$1{\alpha}$, IL-2, IL-4, IL-6, IL-10, and INF-${\alpha}$ showed the same expression pattern. All cytokines were suppressed, except IL-6. These findings indicate that changes in cytokine expression could be used to understand the mechanism of polymicrobial infection in PRDC.
To understand the cytotoxic activity of Machilus thunbergii, which has been used as a traditional oriental medicine, the mechanism underlying the cytotoxic effect of its extract on human acute Jurkat T cells was investigated. The methanol extract of roots (3 kg) of M. thunbergii was evaporated, dissolved in, and then extracted by water. The water-extracted active substance was designated MTWE. When Jurkat T cells were treated with MTWE at concentrations of 0, 25, 50, and $100{\mu}g/ml$, the apoptotic phenomenon of cells accompanying several subsequent biochemical reactions, such as mitochondrial cytochrome c release, activation of caspase-3, and ICAD degradation, was detected in the Jurkat T cells. Moverover. the expression of Bcl-xL, which is a suppressor for mitochondrial cytochrome c release pathway, was reduced in the Jurkat T cells. As DUSP6, a growth suppressor of cancer cells, ranged from 0, 25, 50, $100{\mu}g/ml$ of MTWE, the expression level was elevated in the Jurkat T cells. The apoptotic morphological change of the nuclei was observed by DAPI staining. Although the potential involvement of the other factors and DUSP6 is currently being investigated in more detail, these findings support the notion that MTWE is able to achieve the apoptosis of Jurkat T cells, and it seems that MTWE is useful as a method of evaluating a chemotherapeutic agent or tonic materials for human acute leukemia.
This study was executed to elucidate the degree of satisfaction with working environment and the factors affecting it for workers who are engaged in the composting plants with livestock manure in the Han-river watershed. As an evaluation on site, we applied the self-administered questionaire including general characteristics, working conditions, degree of satisfaction with working environment, and degree of satisfaction with working condition. Simultaneously, we analysed the exposure levels of odor (ammonia, hydrogen sulfide, and odor intensity) in the boundary area of plants. As a result of the study, health condition, odor, work load, dust, and smoking were proved to be the main factors affecting the degree of satisfaction with working environment for workers who are employed in the composting plants with livestock manure in the Han-river watershed. Of them, it was investigated that health condition, odor, and work load accounted for 61.3% of the total degree of satisfaction with working environment and health condition(p$<$0.01), odor(p$<$0.05), work load(p$<$0.05), and dust(p$<$0.05) were statistically significant. It was concluded that the degree of satisfaction with working environment was high for the workers who had good health status and low for the workers who had the relatively considerable work load and were often exposed to odor and dust. Therefore, to increase the degree of satisfaction with working environment for workers who are engaged in the composting plants with livestock manure, the measures that can reduce the exposure levels of odor and dust emitting from the composting plant with livestock manure should be improved.
A total of 96 crossbred weanling barrows aged 21 days were randomly allocated to 32 pens of a new nursery to investigate the effects of antibiotics, phytogenics, and probiotics on intestinal growth and development. The animals were fed a set of three-phase basal diets containing 0.3% zinc oxide (CON) or the basal diets supplemented with 353 ppm of a combination of tiamulin, neomycin, chlortetracycline, and oxytetracycline (ANTI), 75 ppm triterpenoid saponin plus 150 ppm mixed saccharides (HERB; Sacchapin$^{(R)}$), or $1{\times}10^7$ brewer's yeasts plus $8{\times}10^7$ spores of each of Bacillus licheniformis and Bacillus subtilis per kilogram feed (PROBIO; Yeasture Plus 2B$^{(R)}$) for five weeks. Thirty-two pigs representing as many pens were slaughtered at the end of the feeding trial, after which morphological measures and digestive enzyme activities of intestinal mucosa were determined. Weight gain and gain:feed of the pigs were not affected by the dietary treatments (TRT) during the overall feeding trial. Total intestinal length was greater in PROBIO than in ANTI (P<0.05). Wet mucosa weight of the duodenum was not affected by TRT. However, jejunal mucosa weight was greater in PROBIO than in any other group sum of mucosa weights of the duodenum and jejunum was greater (P<0.05) in PROBIO than in ANTI and HERB. The height and width of duodenal villus were not affected by TRT, but crypt depth decreased (P<0.05) in response to HERB and PROBIO vs CON. Specific activities of alkaline phosphatase, sucrase, maltase, lactase, and leucine aminopeptidase in the duodenum and jejunum were not changed by TRT. In conclusion, results suggest that the present dietary treatments have no effects on growth performance of weanling pigs and that of PROBIO enhances intestinal growth and development under a clean experimental setting.
Sa, Soo-Jin;Park, Chun-Keun;Kim, In-Cheul;Lee, Seung-Hoon;Kwon, Oh-Sub;Kim, Myung-Jick;Cho, Kyu-Ho;Kim, Du-Wan;So, Kyoung-Min;Cheong, Hee-Tae;Webb, Bob
Journal of Embryo Transfer
/
v.25
no.3
/
pp.171-177
/
2010
Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. The PA/plasmin system has been associated with a number of physiological processes such as fibrinolysis, ovulation and fertilization. Although correlations have been reported between reactive oxygen species (ROS) and oocyte maturation, the relationship between PA activity and ROS is unknown. The present study was undertaken to determine the effects of cumulus cells on PA activity in matured porcine oocytes under xanthine (X)-xanthine oxidase (XO) system. When oocytes were matured under the X-XO system, the proportion of oocytes remaining GV stage was higher (p<0.05) in oocytes without cumulus cells. The incidence of degenerated oocytes was higher (p<0.05) in the X+XO ($11.1{\pm}6.1$ and $21.6{\pm}3.4%$) than in the control group ($2.9{\pm}1.8$ and $4.0{\pm}1.6%$). The proportion of TUNEL-positive oocytes and activity of caspase-3 were higher (p<0.05) in cumulus-free oocytes and oocytes exposed to ROS. Tissue-type plasminogen activator-plasminogen activator inhibitor (tPA-PAI) and tissue-type plasminogen activator (tPA) activity were detected in oocytes that were separated from cumulus-oocytes complexs (COCs) at 44 h of maturation culture, and only tPA was produced in oocytes that were denuded before the onset of maturation culture. On the other hand, the activities of PA were increased (p<0.05) when oocytes were cultured under the X-XO system. The higher activity of tPA was observed in denuded oocytes (DOs) underwent apoptotic changes by oxidative stress. In COCs, however, tPA-PAI as well as tPA activity was detected and apoptotic changes such as DNA cleavage or caspase-3 activation were not observed. These results suggest that tP A may be relevant to apoptotic cell death in porcine oocytes by oxidative stress.
This study was focused on improvement of milk production in Mongolian dairy industry by artificial insemination (AI) technology, supported by ODA of KOICA in Republic of Korea. This program was started in January 2009 and it is in $3^{rd}$ years. This manuscript summarized the data especially on estrus synchronization and pregnancy establishment in dairy cows (Holstein) this year. A total of 81 dairy cows from 4 private farms (38 from Undarmal milk and that of 30, 8 and 5 dairy cows from Onjin (Enkhbayer), Jargalant, and BRM School farms respectively) were synchronized with 5 ml Lutalyse (i.m.) in the dump of dairy cows and then estrus was detected 2 to 3 days after $PGF_{2{\alpha}}$ injection. The synchronized dairy cows were inseminated with 0.5 ml dairy frozen semen by conventional artificial insemination (AI) techniques. Pregnancy was diagnosed about 60 days after AI by palpation method. About 96.3% (78/81) of synchronized cows were responded to single $PGF_{2{\alpha}}$ injection. Total 75 over 78 dairy cows (90.1%) inseminated were diagnosed as pregnant. The estrus induction and pregnancy rates were very effective using Lutalyse injection and conventional AI techniques in Mongolian dairy cow. The present results indicated that AI after estrus induction in Mongolian dairy cows could be applied to dairy breeding technology for improving breeding efficiency and milk production of the country.
This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.
Kim, Hyeon-Ah;Yun, Mi-Young;Song, Hyang-Hee;Cheong, Kwang-Jo;Yoo, Hwa-Seung
Journal of Pharmacopuncture
/
v.13
no.1
/
pp.53-61
/
2010
Purpose : To investigate the effects of the lavender, lemon and eucalyptus oil mixture on the atopy dermatitis skin lesions induced on NC/Nga Mice by dinitrochlorobenzene (DNCB). Material and Method : For this purpose, we fabricated the oil mixture blending three essential oils (lavender, lemon, eucalyptus : ELL) with one carrier oil (jojoba) and apply it on the atopic dermatitis skin lesions of NC/Nga Mice. Atopic dermatitis in NC/Nga Mice was induced by DNCB treatment on the dorsal skin of mice for 8 weeks. The mixture of ratio of each essential oil drop was 1 (eucalyptus) : 2 (lemon) : 2 (lavender) and this mixture was blended with jojoba oil 50ml (0.025%). The ELL-ointment was supplied for 8 weeks. We evaluated the effects of ELL on cell viability of mouse lung fibroblast, clinical skin features and severity, the level of serum Immunoglobulin (Ig) E & Ig G1, Interleukin (IL)-4, IL-13 and Interferon (IFN)-$\gamma$. Results : ELL showed safety on the cell viability of mouse lung fibroblast compared with control group. The cell viability was measured by SRB method. The effects of ELL on clinical skin features and severity in DNCB-induced dermatitis model of NC/Nga mice was significant compared with control group. EEL also showed significant effects on clinical symptom score compared with control group. Serum IgE & IgG1 level and development of atopy dermatitis skin lesions were evaluated. Serum IgE & IgG1 production was significantly down-regulated in EEL group compared with control group. ELL also down-regulated the levels of IL-4 and IL-13, and up-regulated the level of IFN-$\gamma$ compared with control group significantly. Conclusion : ELL was effective on atopy dermatitis by modulating Th2 related factors.
Nebulin is a giant ($600{\sim}900$ kDa), modular sarcomeric protein proposed to regulate the assembly, and to specify the precise lengths of actin filamints in vertebrate skeletal muscles. Recently, There is an evidence that the nebulin also expressed in non muscle tissue, brain and liver. We identified a new isoform of nebulin from adult brain library by PCR screening. It contains two simple-repeats exon 165, 166 and linker-repeats exon $154{\sim}161$ except exon 159. The nebulin modules M160 to M170 (exon 150 to exon 161) has been shown to bind desmin. In mature striated muscle, desmin intermediate filaments surround Z-discs and link individual myofibrils laterally at their Z-discs and to other intracellular structures, including the costameres and the intercalated discs of the sarcolemma, sarcoplasmic reticulum, mitochondria, T-tubules, and nuclei. Therefore, it is an interesting possibility that the differential splice pathways within the linker region of nebulin modify the affinity of nebulin's interaction with desmin. The specific interactions of nebulin and desmin were confirmed in vivo by yeast two hybrid experiments. To verify in the cellular level the interaction between nebulin isoform and desmin, we transfected COS-7 cell with EGFP-tagged nebulin and DsRed-tagged desmin. Based on evidence showing that despite exon 159 was deleted, the new isoform of nebulin was interact with desmin. This suggest that nebulin in brain may interact with another intermediate filament. The conservation of these ligand-binding capacity in brain and skeletal nebulins suggest that nebulins may have conserved roles in brain and skeletal muscle.
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