Light characteristics are very specific in the aquatic environment. Fish vision and different light spectra perception are related to each species’ natural habit. Light is one of the main environmental conditions and can be easily manipulated in artificial rearing settings. Cholecystokinin (CCK) and mucus-secreting goblet cells are the main regulators of digestion. In this study, we established whether the light spectrum (natural condition, full spectrum: green, 520 nm; red, 590 nm, and blue, 480 nm) influences growth performance and digestive activity related to CCK mRNA expression and mucus-secreting goblet cell activity in order to develop a good management protocol and optimal rearing system for the longtooth grouper. For each light spectrum, fish were reared 12 weeks under a flow-through system and fed commercial pellet diets once daily. At the end of the experiment, the final body weights differed among the fish reared under different light spectra. The highest growth performance value was observed in fish reared under the green light condition. On the other hand, the growth performances of fish in the natural and blue light conditions were drastically decreased in last 3 weeks of the experiment. CCK mRNA expression and mucus-secreting goblet cell activity were significantly higher in the fish under green light condition than in the fish under the natural, red, and blue light conditions. Rearing of the longtooth grouper under the green light condition had positive effects on fish growth performance and digestion. We recommend that the appropriate light spectrum for the artificial culture of the longtooth grouper is the green light condition from the perspective of growth performance and the synergistic effects of CCK and mucus-secreting goblet cells. However, longer light treatment periods are needed in future investigations to clarify the effects of light spectrum on the longtooth grouper. Together with the findings of the present study, such studies would result in better understanding of the digestive physiology and contribute to the development of optimal rearing management for commercial production of the longtooth grouper.

Foxi1, a forkhead family of transcription factor, in narrow and clear cells in epididymis is required for male fertility through regulating transcription of vacuolar H+-ATPase. To understand the regulation of Foxi1 gene activation in epididymis, the effects of steroids and their receptor antagonists and testicular factors on the expression of Foxi1 in epididymal segments were examined in mouse. Epididymis were sampled from adult mice following injections of ICI 182,780 (5mg/head, 2 times for 15 days), dexamethasone (DEX, 0.1,1,10ug/kg/day for 5 days) or oral administration of flutamide (FLM, 100mg/kg/day for 10 days). Otherwise, adult mice were orchidectomized (ORX), rested for 2 weeks, and received testosterone propionate(TP, 3mg/kg/day) for 7 days. In addition, adult male mice were subjected to efferent duct ligation (EDL) and epididymis was collected after 15 days. To study estrogen regulation of Foxi1 gene activation via estrogen receptor α (ESR1), Foxi1 expression was examined in ESR1 knock-out mice epididymis. Expression and subcellular localization of Foxi1 was analyzed by realtime RT-PCR and immunohistochemistry. To search transcription factor binding in the mouse Foxi1 gene promoter, in silico analysis was performed using TESS, TFSEARCH, and Gene-Regulation. ICI 182,780 significantly decreased Foxi1 mRNA levels in caput and corpus but increased in cauda epididymis. Foxi1 mRNA levels in caput epididymis of ESR1 KO mice were significantly lower than those of WT mice, but no significantly changed in corpus and cauda epididymis. Taken together, estrogen differentially regulates Foxi1 gene expression in epididymis. In ORX mice, Foxi1 mRNA levels were significantly increased in epididymis, and which was abrogated by TP. Though FLM did not significantly alter the Foxi1 mRNA levels, androgen may affect Foxi1 gene expression in epididymis. DEX significantly decreased Foxi1 mRNA levels in caput and corpus epididymis at 0.1ug/kg/day and in cauda epididymis at 1ug/kg/day, suggesting that glucocorticoid may negatively regulate Foxi1 gene expression. No significant change in Foxi1 mRNA levels was found after EDL. Foxi1 immunoreactivity was found in the nuclei of narrow cells of caput epididymis including initial segment and clear cells of corpus and cauda epididymis. Of note, in ORX mice, Foxi1-positive narrow cells and clear cells were increased, and which was abrogated by TP. In silico analysis revealed the presence of putative binding sequences for ESR1, AR, and GR in the 5’ upstream region from the Foxi1 promoter. In conclusion, the expression of Foxi1 in narrow cells in caput epididymis might be positively regulated by estrogen via ESR1, which was different from estrogen–ESR signaling in clear cells in corpus and cauda epdididymis. Androgen and glucocorticoid may negatively regulate expression of Foxi1 in all epdididymial segments.

Human embryonic stem cells (hESCs) have the potential for use in regenerative medicine and in the field of basic research. Therefore, effective cryopreservation and storage of hESCs are important for preservation of newly established cell line for various purposes. Despite poor survival and slow recovery after thawing, the conventional slow freezing method is most commonly used for cryopreservation of hESCs due to its simplicity and ease of use for freezing a large number of hESCs appropriate to clinical applications. Here we controlled the clump size (Group Ⅰ; 400～450 ㎛, Group Ⅱ; 800～900 ㎛, and Group Ⅲ; 1500～1700 ㎛) of hESCs at 5 days after plating using a glass pipette during cryopreservation in order to obtain a larger amount of hESCs after thawing. Attachment rates differed significantly (P<0.05) in each of the three groups and the average of attachment rate of GroupⅡ was highest in SNUhES4 and H1. In particular, the attachment rate of Group Ⅱ in SNUhES3 showed a significant improvement with ROCK inhibitor Y-27632. These results indicate that clump size and cell-cell adhesions of GroupⅡ are appropriate for cryopreservation compared to the Group Ⅰ and Group Ⅲ. This method increased cell viability and reduced the recovery time leading to various experiments, and therefore has an advantage for use with hESCs like newly established in particular. We demonstrated that use of this effective cryopreservation method with control of the clump size of hESCs can effectively improve the attachment rate and survival of post-thaw hESCs with and without Y-27632.

The development of humanized culture system of human embryonic stem cells (hESCs) hold promise for therapeutic applications. However, conventional culture system contain animal-derived components such as fetal bovine serum and mouse embryonic fibroblasts that bear a risk of transmitting non-human pathogens and incorporation of non-human immunogenic molecules to hESCs. In this study, we developed an efficient xeno-free hESCs culture system using humanized materials, the CELLstartTM, human foreskin feeder and xeno-free medium containing knockOutTM SR XenoFree (XF-medium) without animal-derived material. The hESCs were gradually adapted to the XF-medium; 25:75, 50:50, 75:25 and 100:0. Two karyotypically normal hESC lines, SNUhES4 and H1, were used for the experiments of xeno-free culture condition. The attachment rates at xeno-free culture system were 52.6±12.4%, 67.0±16.6%, 59.0±13.9%, 28.3±2.9% in SNUhES4, 79.3±5.4%, 53.8±20.9%, 69.4 ±6.4%, 59.8±12.6% in H1 and the spontaneous differentiation rates were 42.2±12.7%, 31.4±2.9%, 40.8±14.5%, 55.2±35.5% in SNUhES4, 35.6±8.5%, 36.4±13.5%, 48.4±7.8%, 80.1±6.0% in H1 in the first four passage. Although the attachment rates were low and the spontaneous differentiation rates were high compared to that of conventional system in the early passages using this humanized culture condition, hESCs in this culture condition were found to maintain hESC characterizations; morphology, expression of cell surface markers and stable karyotype. Our results indicate that simplified compositions of humanized culture system can be applicable to the further optimization for a xeno-free culture of hESCs without the loss of pluripotency and contamination from xenogenic sources.

The anandamide signaling plays various roles in directing reproductive processes. Mouse embryos are shown to express high levels of CB1 receptor (CB1R). It has recently been shown that an analog of anandamide induces autophagy-mediated cell death through stimulation of ER stress response in glioma cells. Since adverse effects of high levels of anandamide agonists on embryo development and implantation are well known, we hypothesized that anandamide mediates an autophagic response in embryonic cells as in cancer cells via highly abundant CB1R on embryos. We tested this hypothesis by using a stable anandamide agonist, Methanandamide (MET) in three embryonic cell systems, i.e., mouse embryonic fibroblasts (MEF), trophoblast stem (TS) cells, and preimplantation embryos from mice. RT-PCR, immunofluorescence staining, and Western blot analysis were used to examine the effects of anandamide on autophagy in these systems. In MEF cells, the conversion of LCI to LCII was heightened by methanandamide (MET), and AM251, a selective CB1 antagonist partially reversed the effects of MET. Treating MEF cells with a high level of MET induces clustering of GFP-LC3, seen as large puncta throughout the cytoplasm. At 28 nM concentration, MET also weakly increased LC3II in TS cells. When MET was injected to day 4 pregnant mice, autophagy was increased in blastocysts in utero as demonstrated by the increased number of LC3 puncta. Formation of numerous autophagic vacuoles was also confirmed by electron microscopic observation. In conclusion, this work suggests that the anandamide-CB1 signaling pathway may be one inducer of autophagy in embryonic cells.

Ligand-bound nuclear receptors (NRs) recruit coactivators such as members of the p160 steroid receptor coactivator (SRC) family and cyclic AMP responsive element binding protein (CREB)-binding protein (CBP) to specific enhancer elements and activate target gene transcription. In the present study, we isolated a novel SRC from the sea urchin Strongylocentrotus nudus (SnSRC) by using the ligand-binding domain of retinoid X receptor as a bait in a yeast two-hybrid screening. The SnSRC (1992 amino acids) interacted with several NRs, including sea urchin estrogen receptor-related receptor (ERR), human and masu salmon estrogen receptors (ERα), mouse ERRγ, rat glucocorticoid receptor α, and rat thyroid receptor β. Furthermore, preferential interacting domains for ERα in the SnSRC are located in the central LxxLL motifs, revealed by the truncation and mutagenesis studies. Interestingly transient knockdown of the SnSRC gene in the sea urchin embryo using morpoholino antisense RNA induced abnormal phenotypes at gastrulation stage such as the lack of primary invagitation and exogastrulation. Together, the present study identified a novel steroid receptor coactivator in an invertebrate species sea urchin and demonstrated its pivotal role in sea urchin embryogenesis. It will be helpful to uncover evolutional and functional origin of the vertebrate counterpart and the detailed function of steroid receptor coactivator during early embryogenesis.

Recent genomic evidences from unfractionated embryonic stem cell (ESC) cultures have demonstrated high levels of concomitant activating (H3K4me3) and repressive (H3K27me3) histone methylations, termed “bivalent marks”, at lineage specific gene loci, demonstrating that all cells residing within the cultures are developmentally equipotent. However, this dogma has been challenged, indicating that ESC cultures are heterogeneous, with individual cells displaying dynamic metastability and failed to make a connection with the variations between cell lines, a broad spectrum of differentiation, continuous phenotypic oscillation, and the expression of lineage specific genes in undifferentiated state. Recently, functional in vitro assays via fractionation of ESC cultures based on comparable expression of some phenotypes (c‐KIT, A2B5, SSEA3, Nanog, Rex‐1, IGFR1, and Stella) revealed a plastic gradient of clonogenicity and lineage specification within ESC cultures reflected by the presence of bivalent marks, which are resolved down to activating “monovalent marks”. More interestingly, dynamic heterogeneity represents a conserved feature on both mouse ESCs and human ESCs as being essentially required for self‐renewal and, more importantly, differentiation. However, it is the most substantive obstacle to control and specify ESCs into desirable cell types. Mostly, differentiation from ESCs has been evaluated by measuring the responses of whole EB populations under the specific inducible conditions, making it difficult to identify, which cell populations are dominantly contributing to differentiated progeny from ESCs. Therefore, further identification of novel transcriptional and phenotypic markers may allow for the isolation and enrichment of more promising target cells for stem cell‐based clinical therapy.

This paper attempts to review the validity of metrical evidence for secondary stress in OE (Old English). Initiated by Huguenin (1901), the tradition of reconstructing secondary stress on the basis of meter builds on the isomorphy between language and meter. Even though the prosodic reconstruction from meter has proven to be fairly useful, there are certain properties to be carefully considered in order to argue for the existence of secondary stress in OE. It is argued that verse types, not alliteration, cannot be a reliable source for the reconstruction of secondary stress. Due to the differences with regard to fundamental assumptions on OE meter, the predictions on the placement of secondary stress are not consistent. In addition, most OE metrical systems are not free of inherent circularity between verse types and secondary stress. It is also demonstrated that given the distinct nature of stress assignment, secondary stress in compounds should be distinguished from that in noncompound words. Moreover OE secondary stress cannot be claimed to exist without the precise definition between compounds and noncompound words being properly reflected in the reconstruction process.

We investigated the effects of temperature changes on the oxygen consumption rhythm in Japanese eels, Anguilla japonica, using an automatic intermittent flow respirometer (AIFR). The endogenous rhythm of the oxygen consumption rate (OCR) in the eels (n = 18; 44-74 cm, 145-690 g), freshly collected by bag net from estuaries, was nearly synchronous with the tidal pattern of the estuarine collection site. The magnitude of mean OCR (mOCR) of eels showed variable range of 82.2 - 116.5 ml O2 kg-1 ww h-1 under constant conditions. In case of increasing temperature from 25 to 38℃, the OCR of eels exhibited a gradually increasing trend with a rhythmic pattern until 36℃. Above 36℃, the rhythms of the OCR dampened and the OCR decreased rapidly at around 36 - 37℃. The OCR of the eels exhibited the maximum value at 38℃, and then it sharply decreased. The results suggested that the critical thermal maximum (CTM) regarding the endogenous rhythms of the eels was at around 36 - 37℃ when water temperature increased at 0.5℃/14 h following the acclimation at 25℃. In case of decreasing temperature (0.5℃/14 h) from 25 to 0℃, the OCR of the eels displayed a abrupt decrease up to 23℃, and between at 23 and 20℃, there was an agitation which showed a slight increase in the OCR with a duration of 1-2 days. Below 9℃, the OCR rhythm of the eels showed a constant state regardless of temperature decreasing. These results suggest that the Japanese eel has an upper incipient lethal temperature at 36℃, with a lower thermal limit at 9℃. The biochemical aspects of the eels influenced by water temperature need to be further studied.

Leaf structure is one of the important agronomic traits. A rolled leaf mutant was induced from an ethyl methane sulfonate (EMS)-treated japonica rice, 'Koshihikari'. The rolled leaf mutant showed phenotypes of reduced leaf width and leaf rolling. In addition, several abnormal morphological characteristics were observed, including dwarfism, defected panicle, delayed germination, and lower seed-setting. Microscopic analysis revealed that the number of small veins was decreased and the sizes of adaxial bulliform cells were reduced in the mutant leaves. The genetic study with two F2 populations from the crosses of the rolled leaf mutant with 'Koshihikari' and Milyang23 suggested that the mutant phenotype might be controlled by a single dominant gene.

In order to simulate a free surface flow in a trench channel, a three-dimensional incompressible unsteady Reynolds-averaged Navier-Stokes (RANS) equations are closed with the model. The artificial compressibility (AC) method is used. Because the pressure

Houttuynia cordata is used as a raw material of oriental medicine to acquire antioxidant and anti cancer effect and to cure a heart disease. Mechanochemical technology not only reduces the size of the object but also changes the chemical properties of the object. Extraction of functional materials from the Houttuynia cordata after grinding as a pretreatment using the mechanochemical technology was conducted in this study to investigate the effect of grinding on the yields and antioxidant activities of the extract. Houttuynia cordata was ground by the planetary ball mill and the morphology was analyzed by SEM. Yields of functional materials were increased from 6.2 g in the sample without grinding to 7.0, 7.8, 8.8 g after grinding of 30 minutes, 1 hour and 2 hours, respectively. Nitrite scavenging abilities were increased from 57-77% to 69-86% as a result of mechanochemical pretreatment. Also, DPPH scavenging abilities for the methanol extraction were increased from 10.01-40.29% to 11.01-49.29% as a result of mechanochemical grinding.

A large number of edible seaweeds are consumed by the coastal peoples of Asia. Some of them are used in traditional remedies in many parts of the world. In this study we investigated effects of supplementation with ethyl acetate extracts of the brown alga Eisenia bicyclis (EBE) on rat macrophage to evaluate the possibilities as immune-modulators. Twelve male SD rats were divided into two groups and the treatments were as follows: A, no Eisenia bicyclis extract (EBE) intake and distilled water ; B, oral supplemented with EBE 200 mg/kg. After 5 weeks of supplementation, rats were sacrificed to assess the effect on peritoneal macrophage functions. We showed no increasing effects on tumoricidal activity, phagocytic activity and NO production in macrophages in EBE supplementation group. However, EBE supplementation suppressed NO-iNOS production and p65 translocation into the nucleus in LPS-stimulated macrophages. Overall, these results suggest that the supplementation of EBE might have an anti-inflammatory effects on NO-iNOS production in macrophages throughout the inhibition of NF-kB activation.

In this study we investigated effects of supplementation with ethyl acetate extracts of the brown alga Eisenia bicyclis on innate immune cells to evaluate the possibilities as an immunomoulator in exercise stress. Twenty male SD rats were divided into four groups and the treatments were as follows: A, no Eisenia bicyclis extract (EBE) (200 mg/kg) intake and maintained at rest ; B, no EBE intake and undergoing exercise ; C, EBE intake and undergoing exercise ; D, EBE intake and maintained at rest. After 5 weeks of oral supplementation, rats were undergoing intensive swimming exercises for 2 h and sacrificed to assess the effects on peritoneal macrophages, spleen cells and natural killer (NK) cells. We showed increasing effects on nitric oxide-inducible nitric oxide synthase (NO-iNOS) production by macrophages and no effects of NK tumoricidal activity and suppressive effects on spleen cell proliferation in exercise group. However, EBE supplementation suppressed NO-iNOS production by macrophages and increased NK tumoricidal activity and spleen cell proliferative response to mitogen in exercise group. Overall, these results that EBE supplementation has differential effects on innate immune response and could be useful as sports nutrition.

Although Jungian interpretation of T. S. Eliot has not been very active for the last half century, a number of reasons make C. G. Jung an attractive tool for reading Eliot. First of all, they were contemporaries undergoing the identical moments of history, responding to them in interestingly similar ways. Secondly, they commonly objected to the positivist trend of their times and tried to revive metaphysical and religious visions of the old. Their ultimate concerns lay in transcendental issues, not in the immediate world. Thirdly, they made their main subject matters out of their visions and other non-empirical materials while resorting heavily to mythic and anthropological studies. Resultantly, Eliot’s works are flooded with archetypal figures, especially those of the mother and the anima. Fourthly, they tried to map out the paths to the Ultimate, sharing many parallel motifs in their courses. Eliot’s literary ideas including impersonality, objective correlative, metaphysical conceit, and collage can all be viewed as a means to make possible transcendental experiences. They encourage the enlargement of cognitive power, a pre-condition for contact with the world beyond. In this sense Jung and Eliot were both shamanic figures who strove to offer remedies to the disorders and the maladies they found haunting their times by retrieving the lost connection to the source of human existence. However, despite his rational interest in the ultimate encounter between human and divine, Eliot has his works overflowing with characters, scenes, and motifs suggesting his inclination toward the mother.

The concept of Original Sin is central to Anglo-Catholic and Roman Catholic theology. In both Paul's epistle to the Romans and Article IX of the Church of England's Thirty-Nine Articles, Original Sin is not seen merely as an aspect among others of a Christian life but an unavoidable condition of existence. This belief in the fallen state of humanity and nature presents the Christian poet with particular difficulties and nowhere are these difficulties more in evidence than in the matter of language. T. S. Eliot's embrace of Anglo-Catholicism within Anglicanism put the matter of language at the center of his later work, especially Ash-Wednesday and Four Quartets. If humans are fallen creatures, the language they use must, in some sense, be fallen too. Eliot recognized this dilemma and adopted a number of stylistic devices in his later poetry to convey his sense of the fate of language in a fallen world. These devices include his use of repetition that suggests a kind of stammering, incomplete grammatical structures and punctuation, self-deprecatory statements, moments of self-exposure and confession. Most notably in both Ash-Wednesday and Four Quartets, Eliot cautions his readers not to be beguiled by the beauty of poetry itself. In 'East Coker' he goes so far as to state baldly that the 'poetry does not matter'.

Suspended and sinking particles were collected in austral summer during ODP Leg 119 to the Indian Ocean sector of the Antarctic Ocean. Field work was carried out at four sampling sites in Prydz Bay. Two of these sites were located in the Outer Bay, and two in the Inner Bay. At the four locations, a total of ten deployments of a sediment trap array were made. The concentrations of chlorophylls and their degradation products both in suspended and sinking particulate matter in Prydz Bay were analyzed using HPLC. Chlorophylls a and c were the dominant algal pigments both in suspended and sinking particles. Because of the abundance of fecal pellets at Site 740, the mean fluxes at 200 m averaged 6 fold greater than that at 50 m. This implies that a dense swarm of zooplankters, presumably large copepods and/or salps, may "feed and excrete" mainly in between 100-200 m depths at this site, closest to land in Prydz Bay. Interestingly, The flux of phaeophorbide a was generally similar in magnitude to that of chlorophyll a throughout the study areas. This is an evidence that materials escaping from near-surface regions in austral summer derive mainly from the gazing of zooplankters. "New production" from sediment-trapped CHL pigment fluxes in Prydz Bay was estimated using f-ratio of 0.15, ranging from 520 to 1,605 μgC m-2 day-1.