Autophagy is a homeostatic degradation process that is involved in tumor development and normal development. Autophagy is induced in cancer cells in response to chemotherapeutic agents, and inhibition of autophagy results in enhanced cancer cell death or survival. Chloroquine (CQ), an anti-malarial drug, is a lysosomotropic agent and is currently used as a potential anticancer agent as well as an autophagy inhibitor. Here, we evaluate the characteristics of these dual activities of CQ using human colorectal cancer cell line HCT15. The results show that CQ inhibited cell viability in doseand time-dependent manner in the range between 20 to 80 uM, while CQ did not show any antiproliferative activity at 5 and 10 uM. Cotreatment of CQ with antitumor agent NVP-BEZ235, a dual inhibitor of PI3K/mTOR, rescued the cell viability at low concentrations meaning that CQ acted as an autophagy inhibitor, but CQ induced the lethal effect at high concentrations. Acridine orange staining revealed that CQ at high doses induced lysosomal membrane permeabilization (LMP). High doses of CQ produced cellular reactive oxygen species (ROS) and cotreatment of antioxidants, such as NAC and trolox, with high doses of CQ rescued the cell viability. These results suggest that CQ may exert its dual activities, as autophagy inhibitor or LMP inducer, in concentration-dependent manner.

최근 전력 전송을 위해 지하에 건설되는 전력구 구조물이 증가함에 따라, 이러한 구조물의 수명 연장은 매우 중요한 문제로 대두되고 있다. 현재까지의 현장 및 실험결과들은 콘크리트 내부의 철근이 콘크리트 피복의 탄산화 현상에 의해 부식될 수 있음을 보이고 있으며,이러한 탄산화에 의한 철근의 부식은 구조물 주변의 높은 이산화탄소 농도에 의해 빈번히 발생할 가능성을 내포하고 있다. 따라서, 본 연구에서는 실제 전력구 현장에서의 철근 깊이와 탄산화 깊이를 측정한 결과를 바탕으로 우리나라의 전력구 콘크리트 구조물에 대한 탄산화위험도를 평가하였다. 현장 데이터를 기반으로 철근 주변에서의 탄산화에 의한 전력구의 사용수명을 평가하였으며, 이를 위해 확률론적 방법인 몬테카를로 기법을 적용하였다. 또한 균열을 유발한 시험체에 대한 탄산화 촉진 실험을 수행하여, 그 실험결과를 바탕으로 균열을 고려한 경우의 전력구의 사용수명을 수치적으로 평가하고 분석하였다.

The aim of this study is to evaluate the possibility of improvement quality of a city inhabitancy and comfort creation in a daily life in an environment of population in developing cities by reconstruction an existing engineering - architectural building and a construction and creations of new intellectual architectural projects.

Guppy has become a model organism for studying behavioral traits such as courtship and mate choice, as well as for understanding ecogeographic adaptation. Unfortunately, studying the early development of live bearers is more complicated than that of oviparous species, due to the inaccessibility of developing embryos for experimental manipulation. Ulrike et al. showed that the embryos could not be cultured for the entire period of their embryonic development. To optimize conditions embryo in vitro culture we established system for varying the concentration of fetal bovine serum in the medium impact on the embryonic development of in vitro embryos. For in vitro culture, embryos were incubated in 8 ml of sterile embryo medium (L-15 [Leibovitz] medium, supplemented with 5, 10, 15, and 20% fetal bovine serum respectively, 20 units/ml penicillin, and 200 mg/ml streptomycin) in a dark incubator at 25℃. Our study found that in 5% of FBS of the medium, embryos can be maintained until the middle-eyed. In 10% and 15% embryos can be maintained constant development; some of them can be fed; however, in 15% it is faster than 10%. And although in 20% of FBS can sustain rapid development of early stage, but ultimately died. According to our experimental data, both 10% and 15% FBS in medium can be used for in vitro culture, the slowly development in 10% FBS appears to be more conducive to observation.

Development of the central nervous system (CNS) occurs normally in mammalian fetus despite lower temperature in the brain region than in the heart. To investigate the effects of temperature niche on the neural differentiation of stem cells in vitro, P19 embryonic carcinoma (EC) stem cells and N2a neuroblastoma stem cells were induced to undergo neural differentiation by retinoic acid and LiCl, respectively. The cells were analyzed for the expression of neural marker genes during 12 days differentiation. Although there were Map2 and NCAM expressions in both groups, no clear difference was found. Similarly, expression patterns of Tuj1 and NF-M were not different in both groups, showing more intensive staining patterns at day 12 than those at days 4 and 8, respectively. However, more cells expressed GFAP markedly at day 12 in 37℃ group. There was little expression of the above markers in N2a cells during differentiation except for Ngn2 and Tuj1. It was found that Ngn2 was expressed more intensely at days 6 and 9 in 33℃ group. Tuj1 expression showed a similar pattern to those of P19 EC cells. RT-PCR analysis also showed that the expressed transcripts did not quite different in both groups, although they were different among the days of differentiation. Thus, it appears that neural differentiation occurs normally with a slight delay and probably less cell death in the cells at 33℃ than that at 37℃.

Chemoresistance is one of the main problems to treat different kinds of cancers or cancer cells. Therefore, it is necessary to find out the strategies to make the cancer cells sensitive to chemotherapy along with optimal dosage of drugs. We examined sensitivity of MCF7 cells through pretreating with an epigenetic modulator, azacytidine (AzaC) to doxorubicin (Dox). The cells were treated with 5 and 10 mM of AzaC for a week, subsequently with 50, 100 and 500 nM of doxorubicin for 24 and 48h. It was found that pretreatment of AzaC significantly enhance the sensitivity of MCF7 cells to Dox, inducing cell death. After 24h 15% cells underwent apoptosis in 500 nM dox treatment group while 23.4% cells death occurred in AzaC pre treatment group. After 48h MCF7 cells treated with Dox showed 19.0% cell death while AzaC sensitized cells showed 50.0% cells death when exposed to 500 nM of Dox for 48h. Western blot analysis showed the upregulations in the expression of bax, caspase-3, caspase-9 and p53 in AzaC-sensitized MCF7 cells treated with Dox as compared to those treated with only Dox. There was no clear indication for pro-apoptosis genes in the cells treated with individual drugs. These results showed that pretreatment with the epigenetic modulator significantly increased the sensitivity of MCF7 cells to Dox. Therefore it is concluded that demethylation event might enhance the activity of DNA intercalating agents to induce DNA damage in breast cancer cells.

Reactive oxygen species (ROS) are produced in organisms as the natural products of oxidative metabolism by environmental stress and pathogen invasion. ROS, such as superoxide anion and hydrogen peroxide, can be toxic to cells and tissues to cause oxidative stress. Recent study revealed that olive flounder (Paralichthys olivaceus) superoxide dismutase (SOD) has been identified as a partial gene and strongly induced to benzoin[a]pyrene and it was deduced indicator of aquatic oxidative stress responses, but its transcriptional response against viral infection has not been investigated. In the present study, spatial and temporal expression profile was analyzed to investigate the function of Of-SOD in the anti-viral response. Of-SOD transcripts were ubiquitously detected in diverse tissues with variable levels using a real-time PCR. The expression of Of-SOD was significantly higher in the muscle, liver and brain, but extremely low in the stomach and spleen. Following VHSV challenge, the expression of Of-SOD increased within 3 hours and subsequently decreased to the original level at 2 days post-challenge in kidney. Although expression pattern and induction time are slight differences depending on the tissue, the transcript of Of-SOD was consistently increased in acute infection response, but expression is low in the chronic response. Collectively, Of-SOD expressions were inducible after VHSV infection and they were probably involved in the immune response against viral challenge. These results suggest that SODs may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in this flounder.

Molecular markers are useful for selecting to include superior character genetic like as strong immune system and rapid growth in fish. The marker is also very important part of breeding technology in Olive flounder (Paralichthys olivaceus). Single nucleotide polymorphisms (SNPs) marker is already in use widely for genomic research and breeding. But this SNPs marker hardly has been validated for screening functional genes in Olive flounder. We study identify single nucleotide polymorphisms (SNPs) on Expressed sequence tag (EST) database, develop usable SNP marker and apply to wild sample and cultured of olive flounder. As a result, Out of total 4.327 ESTs, 693contigs and 514 SNP from total contigs were detected while these substitutions include 297 transitions and 217 transversions. 144 developed markers were applied in 16 samples (wild 8, culture 8), Out of total marker, only 32 markers had detected polymorphic in sample. Polymorphism of 32 markers was observed in the variety genes region involved in immunity and protein synthesis. And the 32 marker were identified 21 transitions, 11 transversions, and indel was not detected in polymorphic SNPs. The analysis on heterozygosity by sample showed 0.34 in wild sample and 0.29 in cultured sample. In conclusion, we was identified SNP and Polymorphism by designed new marker, it supports that development marker is suitable for SNP detection and diversity analysis in Olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Melatonin has several known physiological functions, the main one being synchronization of daily and seasonal rhythms. In addition, melatonin has been reported to influence reproduction and behavioral rhythms with varying results depending on the species. To date, it remains unknown how this rhythm in locomotor activity is controlled endogenously, although there must be coordination of chemical and molecular drivers. However, the species is poorly characterized at molecular level with little sequence information available in public databases. The aim of study was to clarify involvement of endogenous melatonin rhythms and locomotor activity in day-night activity of the eel, Anguilla japonica which is an economically important but endangered species. The levels during daytime (zeitgeber time; ZT 6) were significantly (P<0.05) lower than those during nighttime (ZT 18). A similar pattern was persisted under DD conditions, whereas it disappeared under LL conditions and ocular melatonin levels remained low. Therefore, it is likely that ocular melatonin levels of the nocturnal eel reared under LD and DD conditions fluctuate in a daily/circadian manner and night-related physiological processes are dependent on eel locomotor activities which is a nocturnal species. We found that similar number of genes were differentially expressed between day (ZT6) and nighttime (ZT18), suggesting that during the nighttime also important in differential gene expression with daytime. This work also provides essential information for further studies investigating the molecular basis of daily/circadian system in this species.

As a preliminary investigation into the effect of environmental factors control for gonadal development, we examined the involvement of photoperiod and water temperature in ovarian development of Epinephelus. akaara. For the induction of sexual maturation, E. akaara reared in recirculating aquaculture system (RAS). During November 2013, the photoperiod and water temperature was adjusted to 12L:12D and 18℃, respectively. In the photo-thermal treatment group, every 3 weeks daylight was increased as follows a 13L:11D and 14L:10D, and control group was maintained under natural condition. After 9 weeks, water temperature was increased 23℃ in photo-thermal treatment group. The sampled fish every 3 weeks revealed increase in gonadosomatic index (GSI; 5.18±1.38), oocyte diameter and vitellogenic oocytes (423.9±36.1 ㎛) were observed in gonads 12 weeks under photo-thermal treatment group. However, ovarian development was maintained immature stage in control group. In this environmental factors manipulation trial, seventy one of the 95 females (578.4 ± 25.4 g in mean body weight, 31.0 ± 0.5 cm mean total length) treated with HCG injection (doses 500 IU/kg BW) were induced ovulation by artificial stripping. The total volume of ovulated eggs were 3,470 ml and the total volume of fertilized eggs was 3,295 ml. The fertilization rate and hatching rate were 95% and 98%, respectively. These results suggest that the photoperiod as well as water temperature are major environmental factors in triggering the gonadal development of E. akaara.

Aromatase is an enzyme that converts testosterone to estrogen. This enzyme, present in the sperm as well as various tissue and cells, has been considered to be related to the fertility of human and mouse sperm. Therefore, we examined effect of aromatase inhibitor on viability and fertility of sperm, and quantity of aromatase in sperm groups with different density in pig. To analyze the effect of aromatase on sperm viability, we treated aromatase inhibitor to the sperm with different concentrations (0, 10, 20, 50, 100, 200, 500 μM) at different time (0.5, 1, 2, 4, 8 hours). After the treatment, the sperm viability was calculated by hypo-osmotic swelling test. We selected 0, 50, 100 μM concentration during 0.5 hour as inhibitor treatment condition before in vitro fertilization. Next, we examined fertility and quantified aromatase protein in sperms with different density. In the first experiment, viability of sperm was decreased following the increasement of inhibitor concentration. The aromatase inhibited sperm showed lower penetration rate and cleavage rate than those of non-treated sperm. Concentration of 50 μM inhibitor had no significant effect on the sperm viability, but it significantly reduced sperm fertility. Second, sperms with low density showed higher penetration rate, but no significant difference between sperms with high density. In conclusion, aromatase is responsible for viability and fertility of porcine sperm similar to mouse and human, however, density of sperm has no correlation with quantity of aromatase protein.

We investigated the change mRNA expression of GtHs subunits (FSHβ, LHβ) in the pituitary, androgen receptor (AR), estrogen receptor (ERα) in gonad and histological observation of gonads in longthooth grouper Epinephelus bruneus by treatment Femara, an aromatase inhibitor (AI). Longtooth grouper (body weight 408±43.1 g; one year) cultured in Future Aquaculture Research Center, NFRDI were used in the experiments. The experiment was conducted for 12 weeks from 21 August 2013. Fish received intramuscular injection of AI at 5 mg/g BW dose in three times every 3 weeks. Fish were sampled pituitary and gonads at 3, 6, 12 weeks post-injection (n=50). The mRNA levels of FSH-β, LH-β in pituitary and AR, ERα mRNA in gonad were evaluated using qRT-PCR and qPCR. The histological change of gonads observed on light microscope. The gonads of control group contained most perinucleolus oocyte. At 3 to 6 weeks post-injection, the gonads of AI-treated group contained a few degenerated oocytes, spermatogonia and spermatocytes. At 12 weeks post-injection, gonads contained spermatids undergoing spermatogenesis. From 6 to 12 weeks post-injection, the expression level of GtHs subunits mRNA in pituitary was significantly higher than control group. The expression level of AR mRNA in gonad was higher than control group from 3 to 12 weeks post-injection. The expression level of ERα mRNA in gonad was lower than control group from 6 to 12 weeks post-injection. These results suggest that immature longtooth grouper with AI treatment induced masculinization via change of GtH subunits in pituitary, AR and ERα mRNA in gonad.

Pluripotent cells are categorized as either "naive" or "primed" based upon their pluripotent status. According to previous studies, embryonic stem cells and embryonic germ cells are identified as naive pluripotent stem cells and epiblast stem cells are identified as primed pluripotent stem cells. In a permissive species such as the mouse, naive and primed pluripotent stem cells can be derived from embryos without genetic manipulations. In non-permissive species such as humans and pigs, primed pluripotent cells are only established from embryos. However, previous studies have shown that the embryonic germ cells of non-permissive species share similar morphology and features with naive pluripotent cells. For these reasons porcine embryonic germ cells (pEGCs) may provide a useful cell source for comparative studies on naive pluripotent cells in non-permissive species. In this study, we attempted to establish and characterize porcine embryonic germ cells. Consequently, an embryonic germ cell line was derived from the genital ridges of a porcine dpc 30 fetus in media containing LIF and bFGF. After establishment, this cells were cultured and stabilized in LIF or bFGF contained media. This cell lines displayed a dome-shaped colony morphology in both culture condition. The cell lines were maintained in both condition over an extended time period and were able to differentiate into the three germ layers in vitro. Interestingly, cell lines cultured in LIF or bFGF expressed different pluripotency markers. LIF-dependent pEGCs expressed naive-pluripotency markers such as OCT4, SOX2, NANOG and SSEA1, while bFGF-dependent pEGCs expressed primed-pluripotency markers such as OCT4, SOX2, NANOG and SSEA4. However, as a result of analysis of XCI, two cell lines showed hemi-methylated pattern similarly in XIST promoter regions. In conclusion, we were able to successfully derive embryonic germ cells from genital ridges of a porcine fetus. Pluripotent state of pEGCs were regulated by modulation of culture condition. In LIF supplement, pEGCs showed naive-pluripotency expressing SSEA1, while pEGCs show primed-pluripotency expressing SSEA4 in bFGF condition. This cell line could potentially be used as naive pluripotent cell source for comparative study with porcine embryonic stem cells and other pluripotent cell lines. As porcine pluripotent cells, pEGCs could be useful candidates for preliminary studies of human disease as well as a source for generating transgenic animals.

The paper deals with the impact of the product distribution and information technology sectors on energy resource use, carbon emissions and economic growth by examining the long-run equilibrium relationships and Granger causal relationships among these variables in South Korea. The quarterly time series data from the first quarter of 1970 to the third quarter of 2010 (163 observations) are collected and retrieved from the Bank of Korea database. The paper examines the long-run equilibrium relationships using cointegration techniques and Granger causality using vector error correction models. Test results indicate a long-run equilibrium relationship exists among these variables. In testing directional causality, both the product distribution and the information technology sectors show direct effects on economic growth but only marginal effects on carbon emissions.

Cell cycle process is regulated by a number of protein kinases and among them, serine/threonine kinases carry phosphate group from ATP to substrates. The most important three kinase families are Cyclin-dependent kinase (Cdk), Polo-like kinase (Plk), and Aurora kinase. Polo-like kinase family consists of 5 members (Plk1-Plk5) and they are involved in multiple functions in eukaryotic cell division. It regulates a variety of aspects such as, centrosome maturation, checkpoint recovery, spindle assembly, cytokinesis, apoptosis and many other features. Recently, it has been reported that Plks are related to tumor development and over-expressed in many kinds of tumor cells. When injected the anti-Plk antibody into human cells, the cells show aneuploidy, and if inhibit Plks, most of the mitotic cell division does not proceed properly. For that reasons, many inhibitors of Plk have been recently emerged as new target for remedy of the cancer therapeutic research. In this paper, we reviewed briefly the characteristics of Plk families and how Plks work in regulating cell cycles and cancer formation, and the possibilities of Plks as target for cancer therapy.

The flower buds of Syzygium aromaticum (clove) have been used as traditional medicine for the treatment of male sexual disorders in Asian countries. Recently, there are some reports about the effects of the clove on reproductive activities in mammals. Therefore, its effect on testicular function was examined in male golden hamsters whose reproductive activity is inhibited by photoperiod such as winter climate. The male animals were given by daily oral administrations (56 consecutive days) in three doses (4 mg, 20 mg, and 100 mg/kg BW) of the alcoholic extract of the clove. Generally lower dose (4 mg) of the extract continued to keep the reproductive activities of testes. The both middle and high doses (20 mg and 100 mg) of the extract completely inhibited the testicular activity in some animals. Taken together, these results suggest a possible biphasic action of alcoholic extract of Syzygium aromaticum flower bud on testicular function.

This study was investigated spawning behavior, structure of egg masses and egg development in Aplysia kurodai inhabiting the coastal waters of Jeju Island, Korea. The mating and courtship behavior of A. kurodai occurred in the form of unilateral copulating with chain formation. In chain copulation, only the first animal acted as a female; the second and succeeding animals acted as males (sperm donors) to the animals in front and as females to the animals behind. The fertilized eggs were packaged in capsules that are embedded in jelly to form a cylindrical string called an egg masses. The number of capsule per cm of the egg masses was 55 to 60 capsules and each capsule within the egg masses held 15 to 25 eggs. After spawning, the egg masses were bright yellow or orange in color. This egg masses color not changed until embryos developed into trochophore stage. Thereafter, as embryo developed from trochophore stage to veliger stage the egg masses color became brownish. The fertilized eggs were spherical, with a diameter of approximately 80±1 μm at spawning. At 5 to 6 days after spawning, the embryo developed into trochophore stage and began to rotate within the egg capsule. In the trochophore stage, the precursor of the velum, called the prototroch or prevelum, developed. At 10 days after spawning, the prevelum is transformed into the velum, and the trochophore developed into veliger stage. Between 10 to 15 days after spawning, the veligers broke out of the egg capsule, and hatched as free-swimming larvae.

Science in the 21st century does not consider participants’ welfare, safety and human rights in clinical studies, but modern science puts economic profits in its priority. This leads to a growing concern about social responsibility and professionalism ethics of companies, sponsors and scientists. Specifically, there is no way to control conflicts of participants’ welfare with economic profits, leading to simply relying on individual ethics, social responsibilities and audit. This paper helps relevant agencies and people involved understand conflict of interest. Also this study presents the guidelines as well as independence, autonomy, ethical imagination and phronesis required for scientists.

In mammals, puberty is a process of acquiring reproductive competence, triggering by activation of hypothalamic kisspeptin (KiSS)-gonadotropin releasing hormone (GnRH) neuronal circuit. During peripubertal period, not only the external genitalia but the internal reproductive organs have to be matured in response to the hormonal signals from hypothalamic-pituitary-gonadal (H-P-G) axis. In the present study, we evaluated the maturation of male rat accessory sex organs during the peripubertal period using tissue weight measurement, histological analysis and RT-PCR assay. Male rats were sacrificed at 25, 30, 35, 40, 45, 50, and 70 postnatal days (PND). The rat accessory sex organs exhibited differential growth patterns compared to those of non-reproductive organs. The growth rate of the accessory sex organs were much higher than the those of non-reproductive organs. Also, the growth spurts occurred differentially even among the accessory sex organs; the order of prepubertal organ growth spurts is testis = epididymis > seminal vesicle = prostate. Histological study revealed that the presence of sperms in seminiferous tubules and epididymal ducts at day 50, indicating the puberty onset. The number of duct and the volume of duct in epididymis and prostate were inversely correlated during the experimental period. Our RT-PCR revealed that the levels of hypothalamic GnRH transcript were increased significantly on PND 40, suggesting the activation of hypothalamic GnRH pulse-generator before puberty onset. Studies on the peripubertal male accessory sex organs will provide useful references on the growth regulation mechanism which is differentially regulated during the period in androgen-sensitive organs. The detailed references will render easier development of endocrine disruption assay.

Previous studies, including our own, have demonstrated that the intratesticular injection of hypertonic saline (20%) decreased serum testosterone level which was similar to the surgical castration in the rat, showing the state of chemical castration. In the present study, we further verify the efficacy of this less invasive method as an alternative of surgical orchidectomy in the andrological field. Sterilized 20% saline was directly injected into the adult male rats (750 μl per testis). The tested rats were divided into 3 groups including intact group (intact), orchidectomy group (ORX) and saline injection group (SAL) after bilateral orchidectomy was performed at the same day of injection. All rats were sacrificed at 4 weeks after injection. The reproductive organs (testes, epididymis, seminal vesicles and prostates) were collected and used for DNA and protein pattern analyses. Also, patho-histological studies on the testes were performed. In contrast to the intact group, similar DNA damages of testis and seminal vesicle were appeared in ORX group and SAL group. The DNA degradations seemed to be the results of necrosis rather than apoptosis. In the protein pattern analysis, all the testing tissues exerted similar patterns in the ORX group and the SAL group compared to the those of intact group. Patho-histological studies revealed that severe degenerative changes in testicular seminiferous tubules and massive infiltration of immune cells in SAL group. The present study confirmed that direct injection of hypertonic saline into the testis caused the equivalent biochemical changes in the accessory sex organs as shown in the orchidectomized animals. These results suggest that hypertonic saline injection model could be a useful castration model which can substitute for surgical castration when its safety is secured through further study in the future.