Spermatogonial stem cells (SSCs; also known as Asingle [As] spermatogonia in mice) divide to self-renew or to produce progenitor cells known as Apaired(Apr) spermatogonia in basal compartment of seminiferous tubules of mammalian testis. These characterized cells are the finally differentiated product of a developmental process referred to as “spermatogenesis.” In the development of SSCs it is critical to maintain a balance between self-renewal and differentiation. because an excess of either process will lead to infertility. these two processes are tightly controlled by intrinsic signals of SSCs and extrinsic signals from the microenvironment, known as the SSC niche. The SSC niche is formed by Sertoli cells, the only somatic cells found inside the seminiferous tubules. The WNT/β-catenin pathway is known to regulate Sertoli cell functions critical to their capacity to support spermatogenesis in the postnatal testis, but The mechanisms and factors of the pathway are not well known. We found a factor TLE3 (Transducin Like Enhancer Of Split 3). The transcriptional co-repressor TLE family is known to function as transcription co-repressors within the context of Wnt signaling by interacting with histone deacetylase HDAC2. We examined the expression level of TLE3 in various mouse tissues. As a result of RT-PCR, TLE3 showed significantly higher expression in testis than that in other tissues. Immunofluorescent analysis revealed that TLE3 and HDAC2 expression are differentially regulated in the mouse testis during postnatal development. In adult testis, TLE3 and HDAC2 were co-expressed in Sertoli cells. TLE3 and HDAC2 protein are also located in nucleus in mouse TM4 Sertoli cells. Taken together, TLE3 may play a role in regulating WNT/β-catenin pathway via interaction with HDAC2 in Sertoli cell. Futher studies are needed to look into factors that regulated by siTLE3 in Sertoli cell and interated with TLE3 in WNT/β-catenin pathway.

To characterize the female or male transcriptome of the Pacific abalone and further increase genomic resources, we sequenced the mRNA of full-length complementary DNA (cDNA) libraries derived from pooled tissues of female and male Haliotis discus hannai by employing the Iso-Seq protocol of the PacBio RSII platform. We successfully assembled whole full-length cDNA sequences and constructed a transcriptome database that included isoform information. After clustering, a total of 15,110 and 12,145 genes that coded for proteins were identified in female and male abalones, respectively. A total of 13,057 putative orthologs were retained from each transcriptome in abalones. Overall Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analyzed in each database showed a similar composition between sexes. In addition, a total of 519 and 391 isoforms were genome-widely identified with at least two isoforms from female and male transcriptome databases. We found that the number of isoforms and their alternatively spliced patterns are variable and sex-dependent. This information represents the first significant contribution to sex-preferential genomic resources of the Pacific abalone. The availability of whole female and male transcriptome database and their isoform information will be useful to improve our understanding of molecular responses and also for the analysis of population dynamics in the Pacific abalone.

Gonadotropin-releasing hormone (GnRH) is a key neuropeptide regulating reproduction in humans and other vertebrates. Here, we evaluated the reproductive control system mediated by GnRH in the Pacific abalone Haliotis discus hannai. We cloned a prepro-GnRH cDNA (Hdh-GnRH) from the pleural-pedal ganglion (PPG) in H. discus hannai, and analyzed its spatiotemporal gene expression pattern. The open reading frame of Hdh-GnRH encodes a protein of 101 amino acids, consisting of a signal peptide, a GnRH dodecapeptide, a cleavage site, and a GnRH-associated peptide. This structure and sequence are highly similar to GnRH-like peptides reported for mollusks and other invertebrates. Quantitative polymerase chain reaction demonstrated that Hdh-GnRH mRNA was more strongly expressed in the ganglions (PPG and cerebral ganglion [CG]) than in other tissues (gonads, gills, intestine, hemocytes, muscle, and mantle) in both sexes. In females, the expression levels of Hdh-GnRH mRNA in the PPG and branchial ganglion (BG) were significantly higher at the ripe and partial spent stages than at the early and late active stages. In males, Hdh-GnRH mRNA levels in the BG showed a significant increase in the partial spent stage. Unexpectedly, Hdh-GnRH levels in the CG were not significantly different among the examined stages in both sexes. These results suggest that Hdh-GnRH mRNA expression profiles in the BG and possibly the PPG are tightly correlated with abalone reproductive activities.

Interleukin-12 (IL12) and IL23 are members of the IL12 family and secreted from antigen presenting cells (APCs) such as dendritic cells and macrophages. IL12 and IL23 are composed of two subunits of a sharing subunit, IL12B, and a unique subunit, IL12A for IL12 and IL23A for IL23. IL12 is involved in induction of T helper (Th) type 1 response, whereas IL23 is associated with the differentiation of naive T cells into Th17 cells. It has shown that IL12, a proinflammatory cytokine, is down-regulated during pregnancy for successful establishment and maintenance pregnancy and increases in plasma levels in women with preeclampsia. IL23 decreases IL12 expression to change the immune microenvironment at the maternal fetal interface in humans and mice. In the present study we determined the expression of IL12 and IL23 and their receptors in the endometrium and placenta during pregnancy. Real-time RT-PCR analysis showed levels of IL12A and IL12B mRNAs in the endometrium were high during early pregnancy, but maintained low during mid- to term pregnancy. During pregnancy, levels of IL12RB1 mRNA in the endometrium showed a biphasic pattern with the highest levels on Days 15 and 60 of pregnancy, while levels of IL12RB2 mRNA did not change. Levels of IL23A and IL23R mRNAs in the endometrium decreased toward term pregnancy. Immunohistochemical analysis showed that IL12A protein was localized specifically to scattered cells in endometrial stroma, but barely detected during mid- to term pregnancy. Conceptuses from early pregnancy expressed IL12, IL23, and their receptors, except IL12RB2, and corioallantoic tissues during mid- to late pregnancy expressed IL12, IL23, and their receptors, but not IL23R. These results showed that IL12 and IL23 and their receptors were expressed at the maternal-conceptus interface, suggesting that IL12 and IL23 may play a key role in regulating maternal immune environment for the establishment and maintenance of pregnancy in pigs.

Bovine mammalian gland has biopotential in therapeutic protein production and could be used as a model in further lactation researches. In this study, we have isolated bovine mammary gland-derived epithelial cells (BMECs) from Korean Holstein dairy cattle and themselves show differential dynamic ability in in-vitro culture. BMECs enables to form lobulo-alveolar structure, express milk production related gene. Functional studies indicated that BMECs secret exogeneous antibacterial fragment-Bovine Lactoferricin B (bLfcin-B), which is inserted in PiggyBac system, and this bioactive fragment inhibits the growths of Escherichia coli and Staphylococcus aureus. These data demonstrated that BMECs open new scope in either bioactive fragment, heading to prevent the spread of mastitis or post-mastitis damage in dairy graze and could be an ideal bioreactor for antibacterial proteins.

S100 protein family is small calcium-binding proteins with two EF-hand motifs and comprises more than 20 proteins in human. Although S100A proteins are known to play important roles in proinflammatory responses including damage-associated molecular pattern (DAMP) signaling and in the establishment of pregnancy, the expression of S100As have not been determined in the uterine endometrium during the estrous cycle in pigs. Thus, this study was performed to investigate expression and localization of S100A8, S100A9, and S100A12 in the uterine endometrial tissues during the estrous cycle in pigs. Real-time RT-PCR analysis showed that S100A8, S100A9, and S100A12 mRNAs were expressed in the uterine endometrium during the estrous cycle with higher levels on days 15 and 18 of the estrous cycle than the other days of cycle. Immunohistochemistry analysis showed that S100A9 and S100A12 proteins were mainly localized to the immune cells in the uterine endometrium. Especially, S100A9- and S100A12-positive immune cells were detected in the uterine blood vessels on day 15 of the estrous cycle, and also localized to stroma near to luminal epithelium on days 0 and 18 of the estrous cycle. These results showed that S100As were expressed in the uterine endometrium during the estrous cycle in a cyclic stage-specific manner, and these proteins were localized to the immune cells in the endometrium. These suggest that immune cells expressing S100A proteins may be recruited into the endometrium during the estrous cycle and play an important role in regulating endometrial function in pigs.

The Hippo signaling pathway is essential for regulating proliferation, differentiation, and apoptosis in mammalian cells. Hippo signaling pathway exists in most body tissues and organs, where it controls the size of organs and tissues by keeping cell growth in check and promoting cell death as needed. It has been reported that the members of Hippo signaling pathway are highly expressed in mammalian ovaries and uteri. However, the regulatory mechanism of this pathway in the uterus during estrous cycle regulation remains unclear. Serine/Threonine Protein Kinase 4 (STK4, also known as MST1, a homolog of Hippo in Drosophila) is a major factor of Hippo signaling pathway. However STK4 in the mouse uterus has not yet been examined. The purpose of our study was to determine the expression of STK4 during the estrous cycle and regulation by estrogen in the mouse uterus. We found that STK4 was dynamically expressed in uterine endometrium during the estrous cycle. STK4 highly expressed at the estrus, diestrus, and were found to dramatically decrease as it progressed to the proestrus, metestrus stage of uterus during the estrous cycle. Expression of STK4 was dominant in glandular epithelial and luminal epithelial of proestrus, estrus, and diestrus stage, whereas in metestrus stage, expression of gene intensity was faint. Estrogen or estrogen receptor antagonist ICI 182,780 treatment, in ovariectomized mouse uterus, Expression of STK4 and its downstream genes were increased by estrogen. Our results show that the Hippo signaling pathway is estrogen-dependent in the mouse uterus. These informations will give us on sights to understand uterine dynamics during the estrous cycle.

Due to modern trends with postponing child-bearing and getting worse living environment in women, an ovarian aging increased pregnancy failure and other complications with menopause or premature ovarian failure. Although several theories have been suggested such as mitochondrial malfunction, DNA damage/repair/methylation, caloric restriction, studies regarding ovarian aging-related molecular mechanisms for development of therapeutic methods are insufficient so far. Our objective is to determine molecular pathways of ovarian aging that result in pregnancy failure and other complications in women health to develop treatment strategies. This study is consisted of two parts: in Phase I stage, we analyzed distinct gene expression profile between young and aged mouse ovaries, and in Phase II stage several preferentially expressed genes in both ovaries were selected and analyzed their physiological functions and involved molecular networks related to ovarian aging for development of diagnostic markers and therapeutic methods. Ovaries from 10 week and 11 month-old FVB/NJ female mice with synchronized estrus cycle were collected for this study. A half of each ovary was used for RNA preparation and the other half for histological analysis. Using the Illumina HiSeq 2000 System, preferentially expressed genes were identified. Functional annotation database-based gene-set enrichment analyses and Pathway Studio® were employed to evaluate aging-related molecular networks. These findings were confirmed through qRT-PCR and immunohistochemistry. To validate RNA-Seq data, we examined expression patterns of marker genes (Amh, Bmp15 and Nobox) that were wellknown to be decreased in ovarian aging process. In young or aged ovary, preferentially expressed 876 genes were identified and extracellular matrix (ECM; p<0.001) and chromatin/nucleosome-related (p<0.001) protein-coded genes have the majority in these genes by GOTERM analysis. Amongthem, we selected several candidate genes and confirmed their expression profiles by qRT-PCR and immunohistochemistry followed by molecular network analysis. Regarding molecular interactions in these genes, PathwayStudio® was employed to predict aging-involved molecular networks in mouse ovary. Here we report a couple of candidate molecular networks and medicines (chemicals) for targeting these preferentially expressed genes/proteins. Further analyses are scheduled to produce transgenic animal models and with human ovarian tissues/cell lines.

This paper explores the impact of corporate control, measured by ownership structure, on top-executives’ compensation in Japan. According to agency theory, the pay-performance link is expected to be affected by the firm’s ownership structure. Using a sample of 4,411 firm-year observations (401 firms for the 11-years period from 2001 to 2011) for Japanese non-financial firms publicly traded on the first section and second section of the Tokyo Stock Exchange (TSE), this study demonstrates that institutional ownership (both financial and corporate) is negatively related to the level of executives’ compensation. Such finding is in line with efficient monitoring hypothesis which claims that the presence of institutional shareholders provides direct monitoring over managers, limits managerial self-dealing and curves the increase in top-executives pay. On the other hand, the results also show that managerial ownership is positively related to their compensation which supports managerial power theory hypothesis, i.e. management-controlled firms are more likely to extract more compensation from the business than other firms. Overall, this study confirms that corporate control has significant impact on cash compensation paid to Japanese top-executives after controlling the conventional pay-performance relationship.

Spindle dynamics has a critical role in many physiological mechanisms such as genomic integrity and aging. It is also well known that these mechanisms affect oocyte quality. Because the oocyte quality affects fertility and embryo development, many efforts have been conducted to understand the molecular mechanisms that regulate spindle dynamics during oocyte maturation. Here, we show that translationally controlled tumor protein (TCTP) regulates spindle dynamics during oocyte maturation and prevents deterioration of oocyte quality during postovulatory aging. TCTP was expressed and localized at the cytoplasm with strong enrichment at the spindle microtubules during meiosis. Through the knockdown of TCTP, we found that TCTP regulated stability of the polar microtubules. Meanwhile, spindle dynamics were dramatically decreased with reduced TCTP level during the postovulatory oocyte aging both in vitro and in vivo. Knockdown of TCTP accelerated the reduction of spindle dynamics and aging-related quality deterioration of oocytes. Conversely, overexpression of TCTP rescued microtubule dynamics during postovulatory aging of oocytes and prevented aging-induced deterioration of quality. In addition, overexpression of TCTP improved fertilization competency and subsequent embryonic development. Therefore, our results demonstrate that TCTP is a microtubule-associating protein required to regulate spindle dynamics in mouse oocytes and protects oocytes from aging-related quality deterioration.

Banks play a crucial role in bringing stability and economic development through their expected contribution in proper financial resource mobilisation across the economy. Despite the importance, there is little focus in recent literature which provided the empirical evidence how the global financial crisis affect the bank efficiency in Bangladesh. Thus, this paper aims to examine the effect of the global financial crisis and other factors on the efficiency of Bangladesh commercial banks. By employing the Data Envelopment Analysis (DEA) method, we computed the technical efficiency of individual banks operating in the Bangladesh banking sector during 2000 to 2013. The empirical findings indicate that the Bangladesh banking sector has exhibited the highest efficiency level during 2001, while efficiency seems to be at the lowest level during 2010. The study finds that crisis along with bank size, capital adequacy ratio, return on average equity and real interest rate have a significant effect on bank efficiency in Bangladesh. In order to keep the sound financial development of Bangladesh, banks operating in the Bangladesh banking sector have to consider all the potential technologies which could improve their profit efficiency levels, since the main motive of banks is to maximise shareholders’ value or wealth through profit maximisation.

Purpose - Clarifying one of the biggest public Hospital Costs and incomes according to patients’ case mix. It leads to prepare financial information about pubic medical tariffs and hospital operational costs. Research design, data, and methodology - This study calculates the costs both, with and without taking into account capital costs. This holds for comparison of hoteling based on case mix in all medical procedures. The checklists were reviewed and filled by reviewing accounting documents of the hospital, warehouse exclusion list, and daily books of laundry and CSR. Data was analyzed descriptively by using Excel. Results - In both cases, the hospital is losing in terms of hoteling. Because the buildings and equipment are new, this loss is not tangible. However, this will be revealed when costs of reconstruction and replacement of equipment. The loss rate per day of hospitalization was 569318 Rials for Coronary Care Unit (CCU), 528171 Rials for Post Intensive Care Unit (Post ICU), 474570 Rials for ICU, 233183 Rials for Post CCU and 204803 for Surgical ward. Conclusions - Income of hoteling was lower than its costs. ANOVA showed a strong relationship between case mix and hospital costs as well as case mix and its income. This suggests that optimal case mix can minimize the costs and maximize income.

Little millet (Panicum sumatrense) is well known for its salt and drought stress tolerance and high nutritional value, but very limited knowledge of genetic variation and genomic information is available. In this study, a total of 779 primer pairs were designed from the 22,961 EST sequences of switchgrass (Pancium virgatum), of which 48 EST-SSR markers were developed based on the trials of transferability of these primers in little millet. The EST-SSR amplicons showed reproducible single band polymorphism and produced a total of 160 alleles with an average of 3.3 alleles per locus in 37 accessions of little millet. The average values of expected and observed heterozygosities were 0.266 and 0.123, respectively. The polymorphic information content (PIC) values were observed in range of 0.026 to 0.549 with an average of 0.240. The genetic relatedness among the little millet accessions was evaluated by neighbor-joining dendrogram, which grouped all accessions into two distinct groups. The validation thus demonstrated the utility of the switchgrass EST-SSR markers in assessing genomic relationships in little millet. The findings from this study could be useful for designing strategies for the identification of diverse germplasm for conservation and future molecular breeding programs for little millet.

EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to 1000 ㎍/㎖ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of 50 ㎍/㎖ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of 50 ㎍/㎖. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.

This study analyzed the placement shape and correlation of green space focusing on anion that is one of curing substances. As its result, it was found out that the meteorological element becomes different and the generated amount of anion also gets different along such different meteorological element in case the shape of placing green space. As the result of analyzing the correlation with meteorological element , it was analyzed that it was the positive correlation in case of time, temperature and relative humidity and it was also analyzed that it was negative correlation in case of wind velocity and amount of light. But, in case of wind velocity, it was considered that the further complement research is required as the sampling distribution was shown as so biased by the windproof effect of the trees planted at the outskirts of green space in case of wind velocity. Based on th analyzed results as above, it suggested the model of generating anion by implementing multiple regression analysis such as y = -2462.383 + 0.304x1 x 72.746x2 x 7.315x3 x - 0.138x4 (x1; time, x2; air temperature, x3; relative humidity, x4; luminous intensity, R2; 0.691).

One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-humanprimate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were 127 ± 18.9. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as α-1,3-galactosyltransferase knock-out /hCD46 for xenotransplantation.

This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear maturation and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 subunit beta (Sec61β), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte complexes were collected from follicle with 3-6 mm in diameter. They were incubated in medium with/without EGF for 22 h (IVMⅠ) and subsequently incubated hormone-free medium with/without EGF for 22 h (IVMⅡ). Nuclear maturation state was checked by aceto-orcein stain. Protein expression of EGFR, GM-130, Sec61β, and COPG2 were measured by immunofluorescence. In results, nuclear maturation of oocytes in EGF non-treated oocytes were significantly lower than EGF-treated groups at IVMⅠ or IVMⅡ stage (P<0.05), whereas maturational rate in EGF treatment groups at both of IVM stage was higher in among the all treatment groups (P<0.05). EGFR, GM-130, Sec61β and COPG2 were expressed in the cytoplasm of oocytes. Especially, GM-130 and EGFR were strongly expressed, but Sec61β and COPG2 were weakly expressed in cortical area of cytoplasm. The protein level of GM-130, Sec61β, and COPG2 were significantly higher in the EGF-treated groups (P<0.05). However EGFR was no difference between non EGF-treated groups and control. In conclusion, EGF plays an important role in the systems for oocyte maturation with endoplasmic reticulum and Golgi apparatus. In addition, the protein levels of Sec61β and COPG2 could be changed by EGF in the porcine oocytes during maturation.

The sun is not equally bright over the whole sphere, but rather is darkened toward the limb. This effect is well-known as limb darkening. The limb darkening coefficient is defined by the ratio of the center intensity to limb intensity. In this study, we calculate the limb darkening coefficient using the photospheric intensity estimated from solar images taken by solar and helispheric observatory (SOHO) and solar dynamics observatory (SDO). The photospheric intensity data cover almost two solar cycles from May 1996 to December 2016. The limb darkening coefficient for a size of 0.9 diameter is about 0.69 and this value is consistent with solar limb darkening. The limb darkening coefficient estimated from SOHO shows a temporal increase at solar maximum and a gradual increase since the solar minimum of 2008. The limb darkening coefficient estimated from SDO shows a constant value of about 0.65 and a decreasing trend since 2014. The increase in the coefficient reflects the effect of weakened solar activity. However, the decrease since 2014 is caused by the aging effect.

We present extra-tidal features of spatial configuration of stars around three metal-poor globular clusters (NGC 6266, NGC 6273, NGC 6681) located in the Galactic bulge. The wide-field photometric data were obtained in BVI bands with the MOSAIC II camera at CTIO 4 m Blanco telescope. The derived color-magnitude diagrams (CMDs) contain stars in a total 71´×71´area including a cluster and its surrounding field outside of the tidal radius of the cluster. Applying statistical filtering technique, we minimized the field star contaminations on the obtained cluster CMDs and extracted the cluster members. On the spatial stellar density maps around the target clusters, we found overdensity features beyond the tidal radii of the clusters. We also found that the radial density profiles of the clusters show departures from the best-fit King model for their outer regions which support the overdensity patterns.