• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.24
• /
• pp.10659-10663
• /
• 2015

Background: Rapamycin is an effective anti-angiogenic drug. However, the mode of its action remains unclear. Therefore, in this study, we aimed to elucidate the antitumor mechanism of rapamycin, hypothetically via apoptotic promotion, using MCF-7 breast cancer cells. Materials and Methods: MCF-7 cells were plated at a density of $1{\times}10^5$ cells/well in 6-well plates. After 24h, cells were treated with a series of concentrations of rapamycin while only adding DMEM medium with PEG for the control regiment and grown at $37^{\circ}C$, 5% $CO_2$ and 95% air for 72h. Trypan blue was used to determine the cell viability and proliferation. Untreated and rapamycin-treated MCF-7 cells were also examined for morphological changes with an inverted-phase contrast microscope. Alteration in cell morphology was ascertained, along with a stage in the cell cycle and proliferation. In addition, cytotoxicity testing was performed using normal mouse breast mammary pads. Results: Our results clearly showed that rapamycin exhibited inhibitory activity on MCF-7 cell lines. The $IC_{50}$ value of rapamycin on the MCF-7 cells was determined as $0.4{\mu}g/ml$ (p<0.05). Direct observation by inverted microscopy demonstrated that the MCF-7 cells treated with rapamycin showed characteristic features of apoptosis including cell shrinkage, vascularization and autophagy. Cells underwent early apoptosis up to 24% after 72h. Analysis of the cell cycle showed an increase in the G0G1 phase cell population and a corresponding decrease in the S and G2M phase populations, from 81.5% to 91.3% and 17.3% to 7.9%, respectively. Conclusions: This study demonstrated that rapamycin may potentially act as an anti-cancer agent via the inhibition of growth with some morphological changes of the MCF-7 cancer cells, arrest cell cycle progression at G0/G1 phase and induction of apoptosis in late stage of apoptosis. Further studies are needed to further characterize the mode of action of rapamycin as an anti-cancer agent.

• Bulletin of the Korean Chemical Society
• /
• v.34 no.8
• /
• pp.2419-2424
• /
• 2013

Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

• The Korean Journal of Pharmacology
• /
• v.20 no.2
• /
• pp.23-34
• /
• 1984

To investigate diurnal variations of opiate receptor binding and its modification by experimental condition or treatment of various centrally-acting drugs, the amount of maximum $^3H-morphine$ binding in rat midbrain homogenates was measured at 4 hour intervals for 24 hours. Animals were conditioned under the controlled L : D, 12 : 12 cycle or D: D, 12 : 12 cycle, for 3 weeks and treated with 0.5 ml of physiological saline or drugs for 2 weeks. A highly significant diurnal rhythm with peak at 22 hour of early dark phase with an amplitude$(0.68{\pm}0.06\;pmole/mg\;protein)$ of +51.1% and nadir $(0.33{\pm}0.03\;mole/mg\;prtein)$ at 18 hour of late light phase with an amplitude of -26.6% was found in control group. 24 tour mean of $^3H-morphine$ binding was $0.45{\pm}0.03\;pmole/mg$ protein respectively. Constant dark adaptation or treatment of reserpine, pargyline, imipramine, amphetamine and chlorpromazine modified the diurnal rhythm in the time of peak and nadir binding shape, phase, amplitude of the diurnal curve and 24 hour mean of $^3H-morphine$ binding. However, Kd values were not changed in all experimental groups : Statistical analysis at times of least and great binding indicates that the differences in $^3H-morphine$ binding were due to changes not in the affinity, but in the number of binding sites. The results are interpreted with regard to the diurnal rhythm of opiate receptor finding. The modes of action of psychoactive drugs are closely related to postulated changes of receptor sensitivity in neuropharmacological aspects.

• Tuberculosis and Respiratory Diseases
• /
• v.40 no.6
• /
• pp.659-668
• /
• 1993

Background: p53 is currently considered as a tumor suppressive gene product, and its alterations are suggested to be involved in several human malignancies, including non-small cell lung cancers. p53 expression rates are variable in many reports and among cell types. Also, whether the phase of p53 expression is early or late during carcinogenesis is not certain. Thus, We have investigated to evaluate p53 expression rates of the various cell types and tissues and identify expression phase (early or late). Method: We obtained 71 tissue from 50 non-small cell lung cancer patients and performed the simple immunohistochemical staining using nonspecific monoclonal antibody(NCL-p53DO7). Results: 1) In non-small cell lung cancer patients. the expression rate of lungs(46.5%) is higher than that(25.0%) of lymph nodes. But, there is no significant difference between two groups. 2) Among the various cell types, p53 expression rates in squamous cell carcinoma and adenocarcinoma are 58.3% and 50.0% respectively without significant difference. 3) p53 expression rates in various stages are 33.3%, 60.0%, 40.0%, 60.0% and 66.7% in stage I, II, IIIa, IIIb and IV, respectively with no significant difference. 4) p53 expression rates in the various T parameters are 33.3%, 50.0%, 16.7% and 100% in T1, T2, T3 and T4, respectively and p53 expression rates in the various N parameters are 27.3%, 22.2% and 25.0% in N1, N2 and N3, respectively. There are no significant differences in the expression rates among varous T & N parameters. 5) p53 expression rates of lymph nodes in patients who have positive stains in lungs are 12.5% and 50.0% in N1 and N2. 6) p53 expression rates of all lymph nodes in patients who have negative stains in lungs are 0.0%. Conclusion: The above results show that p53 expression rate in non-small cell lung cancers is not correlated with cell type and progression of stage and it is thought to need further investigations about at what phase p53 expression influences the development and progression of lung cancers.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.12
• /
• pp.77-87
• /
• 1972

The fruiting phase of rape under transplanting and direct-sowing conditions has been studied at Mokpo during the 2 years period from 1970 to 1971. Two varieties, Yudal and Miyuki were used in this study. The planting space and sowing time were also incorporated into this study. The results could be summarized as follows: 1. The plant tape of rape was nearly umbrella-shaped of all, but has changed to the laid elliptical-shaped, broadly ovate and spindle-shaped under different varieties and cultural conditions in the plant diagram(Fig. 2). 2. The length of the primary branches for each nodes had a tendency to the symmetric apical curve with the apex at the upper 10-12th node in the transplanting. but to the upper bias apical curve with the apex at the upper 5-7th node in the dense-sowing(Fig.3). 3. The ear of main stem was longer, more pods, heavier 1, 000 grains and more grain yield than ear of primary branches of all, Especially, as for that, the rate of yield constitution per plant in the direct-sowing was higher than in the transplanting(Fig. 4, 5, 6, 7, 8, 9). 4. The ear-length of the primary branches for each nodes had a tendency to the relatively slowly apical curve with the apex at the upper 3-4th node in the transplanting, but to the lower bias apical curve with the apex at the upper 2nd node in the dense-sowing. Especially, the possibility of growth at the lower ears was few in the early variety (Fig. 4). 5. The number of pod per ear on the primary branches for each nodes had a .tendency to the curve of ear-length with the apex at the upper 5-8thnode in the transplanting and at the upper 4-5th node in the dense-sowing (Fig. 5). Accordingly, a high positive correlation was found between the ear-length and number of pod per ear (Table 2) 6. In the transplanting, the high rate of effective ear was from the upper nods to the 12th node, but below the 16-17th nodes was ineffective. However, in the early dense-sowing the high rate of effective was to the 7th node. but below the 10th nodes was. ineffective. Especially, in the early variety has difficult to secure of poi-numbers for ineffective of the lower nodes(Fig. 6.). 7. The density of pod setting of the ear of main stem was the longest of all ears, and the lower nods were, the shorter it became. That had a tendency to the evidently apical growth. However. in the early variety, it was lengthened according to growth of ear-length(Fig.7). 8. The pod-length of the medium nodes was longer than the upper and lower, and the possitive correlation between pod-length and number of grain per poi was very high(Table 2.). 9. In the grain yield per node of primary branches, the most yielding node of transplanting was the upper 9th node, of dense-sowing 4-5th node(Fig 8.), and the possitive correlation between grain yield per node and ear-length or number of pod per ear was very high(Table 2). 10. The grain yield of ear of main stem was higher than that of primary branches in the percentage of dependence for grain yield per plant. The limint node of 50% of dependence to cumulative grain yield per plant was the upper 7-8th node in tranplanting, in the early dense-sowing 4-5th node, and in tke late dense-sowing-3th node(Fig. 9). 11. In the weight of 1, 000 grains the lower nodes were, the lighter it becames in dense-sowing. Therefore, this was also lighter than in the transplanting to the (Fig. 10.). 12. The oil content of grain at the medium nodes was low in the early variety, but at the ear of main stem and upper 1st node it was extremely high(Fig. 11.).

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.10
• /
• pp.1518-1523
• /
• 2003

This experiment was conducted to investigate the effect of various feeding regimens on growth performance, nutrient digestibilities, nitrogen retention, blood urea nitrogen (BUN) concentration and feed cost in young pigs weaned at 21 days of age. One hundred twenty crossbred pigs (Landrace${\times}$Large White${\times}$Duroc, average 6.8 kg BW), weaned at 21 days of age, were allotted to 5 treatments in a 5 replicates by a randomized completely block (RCB) design. Treatments were as follow: 1) 1P (1-4 weeks: CP 23% and lysine 1.60%), 2) 2P-I (1 week: CP 23% and lysine 1.60%, 2-4 weeks: CP 21% and lysine 1.45%), 3) 2P-II (1-2 weeks: CP 23% and lysine 1.60%, 3-4 weeks: CP 21% and lysine 1.45%), 4) 2P-III (1-3 weeks: CP 23% and lysine 1.60%, 4 week: CP 21% and lysine 1.45%), 5) 3P (1 week: CP 23% and lysine 1.60%, 2-3 weeks: CP 21% and lysine 1.45%, 4 week: CP 19% and lysine 1.30%). Three different diets were formulated and supplied according to phase feeding programs. Diet 1 contains 23% crude protein and 1.60% lysine, diet 2 contains 21% crude protein and 1.45% lysine and diet 3 contains 19% crude protein and 1.30 lysine, respectively. Although there was no significant difference in growth performances, there was a beneficial effect of 3 phase feeding. The ADG was higher in 3P treatment than other treatments and it was observed clearly in late period (3-4 weeks) than in early period. Also, with increase in age, growth rate of pigs in 3P treatment was higher than that in 1P treatment approximately 37% (p=0.1379). There were no significant differences among all treatments in nutrient digestibility. The concentration of BUN was higher in pigs were fed diet containing 21% crude protein and 1.45% lysine (eg, 2P-1 and 3P) than those supplied diet containing high nutrient value at 2 week. The lowest feed cost/kg weight gain of pigs showed in 3P among treatments (p<0.05) whereas, high feed cost/kg weight gain of pigs was calculated in 1P and 2P-II treatments compared with 2P-I and 2 P-II (p<0.05), because of high milk products were used in those diet.

• Journal of the Korea Institute of Building Construction
• /
• v.17 no.2
• /
• pp.191-197
• /
• 2017

The safety nature of construction industry differs from that of manufacturing sector. For instance, accident risk level dramatically varies at each phase of construction process. Korean Occupational and Health Safety Act has been regulated OHS expense and it contributed reducing accident risk and enhancing safety culture for many years. However, although current regulation guides to use OHS expense proportionate to construction progress, it still allows late use of the expense. This study was conducted for the purpose of presenting methods of estimating a step-by-step OHS expense rate required at each construction phase. In order to do provide proper OHS expense schedule, it analysed accident risk of each construction phase by sorting out 1439 cases of construction site fatality reports, and proposed a method of generating appropriate OHS expense scheme according to its construction work progress characteristics. Both linear and sigmoidal S-curve model were used for the analysis, and the latter generally requires earlier use of OHS expense. By comparing the estimated OHS expense use schedule with current criteria, more than 27%p early use of OHS expense is required for the prevention of accident.

• Korean Journal of Poultry Science
• /
• v.45 no.2
• /
• pp.81-88
• /
• 2018

The present study investigated effect of supplementation of agricultural byproduct on growth performance and blood parameters of broiler chicken using meta-analysis. Total 11 studies (32 experiments) where broiler feeding trials with agricultural byproducts were included were employed in analysis. All studies were conducted at commercial farm placed in South Korea and under commercial feeding programs. Growth phases were divided into two groups based on slaughter day. For growth performance, body weight, body weight gain, feed intake and feed conversion rate (feed/gain) were investigated. Carcass weight, dressing percentage, liver and spleen weight, blood glucose, total cholesterol and triglyceride, HDL (High density lipoprotein) and LDL (Low density lipoprotein) cholesterol contents in blood were investigated. Body weight of broiler of treatment was significantly increased than the control (P<0.05), regardless of growth phase. However, body weight gain during late growth phase did not show significant difference between control and treatment. Effect evaluated by random effect model on feed intake was not significant. Effect on feed conversion rate during early growth phase was significantly negative (P<0.05). Effects on carcass weight, dressing percentage, liver and spleen weight were not significant in random effect model. Effect on abdominal fat was significantly negative (P<0.05). In blood parameters, significant effects were found at total cholesterol, triglyceride and HDL cholesterol (P<0.05). Effect on HDL cholesterol only was positive effect, and others were negative.

• Journal of Veterinary Clinics
• /
• v.25 no.6
• /
• pp.488-493
• /
• 2008

This study compared the effect of injectable combinations of anesthetics on each of the fluorescein angiographic phases in order to determine the most useful anesthetic combination for the procedure. Acepromazineketamine (AK), xylazine-ketamine (XK), diazepam-ketamine (DK) and zolazepam-tiletamine (ZT) group were administered randomly to 8 dogs with a two-week interval between different combination doses. The vital signs including the heart rate and arterial pressure were measured before anesthesia and every five minutes during anesthesia. Serial angiographic images were obtained after injecting a sodium fluorescein dye (25 mg/kg) and the onset time of arterial phase (AP), arteriovenous phase (AVP), early venous phase (EVP) and late venous phases (LVP) were recorded. The onset time of the AP, AVP and EVP were significantly slower in the AK and XK groups than in the DK and ZT groups. The total duration of the AP and AVP in the AK group was significantly longer than those in the ZT group. The heart rates were significantly higher in the DK and ZT groups. The arterial pressure was significantly higher in the AK and XK groups (p<0.05). There were significant differences in each angiographic onset time and duration depending on the changes in the heart rates and arterial pressure. The AK and XK groups showed a long angiographic duration allowing an accurate evaluation. Overall, it is believed that AK and XK are more useful for performing fluorescein retinal angiography than DK and ZT.

• Development and Reproduction
• /
• v.16 no.4
• /
• pp.363-370
• /
• 2012

The outer dense fiber 2 (ODF2) protein is an important component of sperm tail outer dense fiber and localizes at the centrosome. It has been reported that the RO072 ES cell derived homozygote knock out of ODF2 results in an embryonic lethal phenotype, and XL169 ES cell derived heterozygote knock out causes severe defects in sperm tail development. The ODF2s splicing variant, Cenexin1, possesses a C-terminal extension, and the phosphorylation of serine 796 residue in an extended C-terminal is responsible for Plk1 binding. Cenexin1 assembles ninein and causes ciliogenesis in early stages of the cell cycle in a Plk1-independent manner. Alternatively, in the late stages of the cell cycle, G2/M phase, Cenexin1 binds to Plk1 and results in proper mitotic progression. In this study, to identify the in vivo function of Plk1 binding to phosphorylated Cenexin1 S796 residue, and to understand the in vivo functional differences between ODF2 and Cenexin1, we generated ODF2/Cenexin1 S796A/Cenexin1 WT expressing transgenic mice in a RO072 ES cell derived $ODF2^{+/-}$ knock out background. We observed a severe defect of sperm tail development by ectopic expression of Cenexin1 S796A mutant and no phenotypic differences between the ectopic expression of ODF2/Cenexin1 WT in $ODF2^{+/-}$ background and in normal wild type mice.