Recently, we have shown that some endocrine disruptors, heavy metals, organotins and azoles suppressed steroidogenic enzymes such as P450 side-chain cleavage enzyme (P450scc) and aromatase in bullfrog ovarian follicles. In the present study, by using an amphibian ovarian follicle culture system, we examined the effects of these endocrine disruptors on maturation and ovulation of oocytes from Rana dybowskii in vitro. Ovarian fragments or isolated follicles were cultured for 24 h in a medium containing frog pituitary homogenate (FPH) or progesterone ($P_{4}$) with or without endocrine disruptors, and oocyte maturation (germinal vesicle breakdown, GVBD) and ovulation were examined. Among the organotins, tributyltin (TBT) strongly inhibited both FPH-and $P_{4}-induced$ oocyte maturation ($ED_{50}$:0.6 and 0.7 ${\mu}M$, respectively); however, tetrabutyltin (TTBT) and dibutyltin (DBT) showed only partial suppression, while monobutyltin (MBT) showed no inhibitory effect. All of the organotins suppressed $P_{4}-induced$ oocyte ovulation very effectively at a low concentration, and TBT and DBT exerted an inhibitory effect on FPH-induced ovulation. Among the heavy metals, mercury (Hg), cadmium (Cd) and cobalt (Co) were very effective in inhibiting FPH-induced oocyte maturation and ovulation, while lead (Pb), arsenite (As) and zinc (Zn) were less effective. However, all of the heavy metals suppressed FPH-induced oocyte ovulation at a high dose ($100{\mu}M$). Among the azoles, itraconazole (ICZ), ketoconazole (KCZ) and clotrimazole (CTZ) effectively inhibited FPH-induced oocyte maturation and ovulation, while econazole (ECZ), miconazole (MCZ) and fluconazole (FCZ) were considerably less effective. These results demonstrated that the abovementioned endocrine disruptors exhibited differential effects on oocyte maturation and ovulation in amphibian follicles and that the frog ovarian culture system could be used as an effective experimental tool to screen and evaluate the toxicity of various endocrine disruptors in vitro.

The naphthoquinone analog (2,3-dichloro-6,9-dihydroxy-1,4-naphtoquinone, NA) has an inhibitory effect on cdc25A protein phosphatase in vitro, which is responsible for G1/S transition during cell cycle. However, the exact mechanism inducing the growth inhibition is not understood. In this study, we investigated the regulatory mechanisms of growth arrest induced by NA, as a new potent inhibitor of cdc25A phosphatase, in human hepatocarcinoma SK-hep-1 cells. We found that NA induced the G1 arrest by perturbation of protein tyrosine dephosphorylation of Cdk2, which may be resulting from inhibition of cdc25A phosphatase. In addition, p21 was expressed in a p53-independent manner and participated in the NA-induced G1 arrest by inhibiting Cdk2 activity. Although the exact mechanism is not known, the p21 expression might be related to MAPK activation. From these results, we suggest that NA induces G1 arrest via inhibition of cdc25A and induction of p53-independent p21 expression in SK-Hep-1 cells.

The effects of antimetabolite 6-AN (6-amino-nicotinamide) on viability and morphology of L6 myoblast cells have been investigated. 6-AN ($100{\mu}M$) induced a time-dependent decrease in cell viability with respect to the untreated control cells. Following 6-AN administration the viability rate started to decline sharply, reaching about 23% of the untreated control cells at 48 h. Inverted phase-contrast microscopy revealed that 6-AN caused characteristic morphological changes such as irregularly elongated and stellate shape of cells, round-shaped nucleus, cytoplasmic vacuolization, irregular cell arrangements and formation of large spaces among cell clusters. The concentrations of ATP and $NAD^{+}$ in the 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. In contrast, the concentration of AMP was significantly increased by the 6-AN treatment. Activities of catalase, superoxide dismutase and glutathione peroxidase in 6-AN treated cells were significantly higher (p < 0.01) than those of the untreated control cells. The activities of glyceraldehyde-3-phosphate dehydrogenase in 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. The results suggest that 6-AN caused marked reduction of cell viability and alterations of some important metabolites and enzymes.

Programmed cell death was characterized in the silkworm thoracic ganglia TG1, TG2 and TG3 during postembryonic periods by TUNEL assay. Apoptotic cells were detected in the three TGs of all larval stages except for day-1, 2 1st instar larvae, in which no apoptotic cells were found. From day-7 5th larva, the numbers of apoptotic cells were dramatically increased and peaked on day-1 pupa and day-2 pupa and then abruptly decreased. Apoptotic cells finally disappeared in day-1 adult. In-vivo injection of 20-hydroxyecdysone (20E) into day-8 5th larva resulted in a striking decrease of apoptotic cells. Actinomycin D (Act D) or cycloheximide (CHX), injected into hemolymph of day-8 5th larva, resulted in a decrease of apoptotic cells in the three TGs. Injection of caspase-8 and -3 inhibitors also blocked cellular apoptosis. These results will provide valuable information for understanding of cellular changes in the three TGs during metamorphosis of the insect species.

Liposome-entrapped atypical Aeromonas hydrophila antigen was prepared to investigate the potential protective efficacy for A. hydrophila infection. Carp (Cyprinus carpio) were immunized orally with liposome-entrapped A. hydrophila antigen. After immunization, significantly more antigen-specific antibodies were detected in serum, intestinal mucus and bile than non-immunized control group. The immunized carp were then challenged by immersion with $1{\times}10^{6}$ cfu/ml of A. hyrdophila for 60 min. Of the eight non-immunized carp, three carp died (62.5% survival), whereas five out of six (83.5%) of the immunized survived. Furthermore, development of skin ulcers was significantly inhibited in carp immunized with liposomes containing A. hydrophila antigen. These results suggest that liposomes containing A. hydrophila antigen have a potential for induction of protective immune responses against atypical A. hydrophila infection and also suggest the possibility of developing a vaccine that may ultimately be used for prevention of fish diseases.

Tau plays a role in numerous neuronal processes, such as vesicle transport, microtubule-plasma membrane interaction and intracellular localization of proteins. SUMO (Small Ubiquitin-like Modifier) modification (SUMOylation) appears to regulate diverse cellular processes including nuclear transport, signal transduction, apoptosis, autophagy, cell cycle control, ubiquitin-dependent degradation, as well as gene transcription. We noticed that putative SUMOylation site is localized at $^{340}K$ of $Tau(^{339}VKSE^{342})$ with the consensus sequence information (${\Phi}KxE$ ; where ${\Phi}$ represents L, I, V or F and x is any amino acid). In this report, we demonstrated that $^{340}K$ of Tau is the SUMOylation site and that a point mutant of Tau S214E (an analog of the phospho $^{214}S$ Tau) promotes its SUMOylation at $^{340}K$ and its nuclear or nuclear vicinity localization, by co-immunoprecipitation and confocal microscopy analysis. Further, we demonstrate that the Tau S214E (neither Tau S214A nor Tau K340R) mutant increases its protein stability. However, the SUMOylation at $^{340}K$ of Tau did not influence cell survival, as determined by FACS analysis. Therefore, our results suggested that the phosphorylation of Tau on $^{214}S$ residue promotes its SUMOylation on $^{340}K$ residue and nuclear vicinity localization, and increases its stability, without influencing cell survival.

The first zoea of Pugettia gracilis is described and illustrated for the first time. Its morphological characteristics are compared with those of other known species of the genus from the northern Pacific waters. Although the Pugettia zoeas of the northwestern and the northeastern Pacifies are very similar, they can be easily distinguished by their dorsal carapace spine. In the northwestern Pacific it is spinulate with a spinous tip, while in the northeastern Pacific it is smooth with a blunt tip.

Four new species, Semitaspongia jejuensis, Scalarispongia regularis, S. nigra and Dysidea violata are described from Jejudo, Korea. They were collected from 15 m depth by SCUBA diving and from intertidal area by hand. Semitaspongia jejuensis n. sp. is easily distinguished from other Semitaspongia species in growth form, conules, colour, skeletal structure, diameter of fibre and habitat. Scalarispongia regularis n. sp. is very close to S. scalaris (Schmidt, 1862) in skeletal structure, but S. scalaris has longer conules, larger meshes, longer distance between primary fibres, and highly developed subdermal canal system. Scalarispongia nigra n. sp. is very close to S. regularis in skeletal structure but can be easily distinguished by its black colour of external surface and growth form. Dysidea violata n. sp. is similar with D. ethria (Laubenfels, 1936) in sponge appearance, but D. etheria is clearly defined by its blue colour and thickness of fibre.

A new cyclopoid species belonging to the genus Acanthocyclops is described from several mountain springs in South Korea. This species is allied to A. kieferi species group in sharing 11-segmented antennules, but is, clearly distinguished from them by its single apical spine on the third endopodal segment of leg 4 and an extra spine on the distal segment of leg 5 in both sexes.

Both sexes of a new species of Miraciidae belonging to the genus Amonardia Lang, 1948 are described. All materials collected from the cultivated brown alga, Undaria pinnatifida of Gijang, Korea. So far only one species, A. normani (Brady, 1872) from the algal bed at Jindo Island was recorded in Korea. The new species can easily be distinguished from its congeners by the combination of characters as follows: (1) shape of first antennular segment in female, (2) setal formular of mandible and maxillule, (3) setae of female sixth leg in female, and (4) shape of P2 endopod and exopod of fifth leg in male.

Eight zoeal stages and one decapodid of the hippolytid shrimp Latreutes acicularis Ortmann, 1890 are described from laboratory-reared material hatched from egg from ovigerous females collected from Shimoda, Japan. The presence of a minute dorsomedian protuberance on the third abdominal somite, the spinules on the dorsal margin of the fourth and fifth abdominal somites, and a pair of dorsolateral spines on the fourth and fifth abdominal somites readily distinguish the larvae of L. acicularis from those of L. anoplonyx Kemp, 1914 and L. laminirostris Ortmann, 1890, the other two known larvae of Latreutes from East Asia.