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Scombrops boops and Scombrops gilberti are commercially important species inhabiting waters around the Japanese archipelago, and recently another undescribed species Scombrops sp. was found. Here, we developed a method using PCR-RFLP on cytochrome b gene sequences of known scombropids to distinguish among these taxa. Comparison of RFLP of the PCR products among scombropids produced a species-specific banding pattern, identifying them with close to 100% accuracy. Two misidentified specimens were identified correctly by the PCR-RFLP analysis developed previously using 16S rRNA sequences. These results showed that PCR-RFLP analysis, using 16S rRNA and cytochrome b sequences in tandem, correctly distinguishes among the three scombropids. We applied this method to 190 individuals collected around the Yonagunijima Island and got some insights into reproductive biology of the three species of Scombrops.

The fish family Scombropidae consists of a single genus, Scombrops, and is distributed in the waters of the northwestern Pacific Ocean, southwestern Indian Ocean and western Atlantic Ocean, including the Caribbean Sea. Among these, the population of South Africa has been renamed from Scombrops dubius to Scombrops boops, which is known to be distributed in waters around Japanese Archipelago. We are skeptical of this revision as the two populations are geographically isolated between the waters of the Far East and the southern end of the Africa coast. Recently, our lab determined the sequences of the complete mitochondrial genome of three Japanese gnomefish, S. boops, Scombrops gilberti and an undescribed scombropid species (Scombrops sp.). Here, we compared the partial sequence of cytochrome c oxidase subunit I (COI) from the three Japanese gnomefish with that of the African population. The African sequences showed 95.5–96.6% identity with the Japanese sequences, while the corresponding sequences from the Japanese species showed 98.2–100% identity with each other. A maximum likelihood tree based on the partial sequence of COI also demonstrated that the clade of African gnomefish is distinct from those of Japanese species including S. boops. These results suggest that the scombropid fish in the African waters is a different species from S. boops and that it is appropriate to resurrect the name Scombrops dubius for the African population.

The early life history of girellid fishes in Japanese waters is unclear, and little is known about their species-specific reproductive strategies. We examined seasonal changes of distribution patterns for settlement-stage juveniles of Girella punctata and Girella leonina on the rocky shore in the regions of Kanto and Izu, Japan, to infer the influence of the Kuroshio Current on their reproduction. We collected 813 settlement-stage juveniles mainly in Sagami Bay and genetically identified the species. The juveniles of G. punctata were collected on the rocky shore in Sagami Bay during April to August, with the abundant catch in May and June. Thus, we infer that juvenile G. punctata ubiquitously inhabit the rocky shore in the area in spring and summer. By contrast, juveniles of G. leonina were rarely collected in Sagami Bay, with a total catch of only 66. Notably, no juveniles were collected during the wintertime in Sagami Bay, although an abundant catch of G. leonina had been previously reported for Sagami Nada off Sagami Bay during January to March. This clear-cut difference between the areas likely reflects the difference in proximity to the path of the Kuroshio Current. We expect that the Kuroshio Current strongly influences the reproductive success of G. leonina.

The feeding ecology of Great Cormorants (Phalacrocorax carbo) during the breeding season in the Kano River basin, central Japan, was examined to clarify the trophic relationship between the cormorants and ayu (Plecoglossus altivelis altivelis) reared for mass release in the river. The ayu was most frequently found in stomachs of cormorants culled during the breeding season, despite relatively poor catch in the year-round fish fauna research in the watershed. Carbon and nitrogen stable isotope ratios of some ayu individuals extracted from the stomachs of the culled cormorants were similar to the isotopic values of ayu caught in the watershed, whereas the other stomach-content ayu showed peculiarly high nitrogen isotopic values, clearly distinct from the values of the ayu caught in the watershed, and overlapped with the values of mass-release ayu. Furthermore, isotopic values of past diets inferred by the isotope analysis of livers of the culled cormorants were closer to the values of the mass-release ayu, relative to the past diet values inferred by the analysis of the cormorant muscles. This suggests that the food supply from the mass-release ayu had increased in the breeding season, since the isotopic turnover rate is faster in livers than in muscles. The huge number of formula-fed ayu released in the watershed create an anthropogenic food chain which is assumed to significantly support the breeding of the cormorants.

Current literature states that family Scombropidae consists of a single genus Scombrops comprising three species worldwide, with two of them, Scombrops boops and Scombrops gilberti, distributed in the waters around the Japanese Archipelago. Although these two scombropids are commercially important species, little is known about the ecology of these fishes. It is difficult to discriminate between these two species based on external characteristics because of their morphological similarity. Here, we report two different morphotypes characterized by the relative growth between the otolith size and the standard length (SL) of the scombropid specimens caught in southern waters off Kyushu Island, Japan, and show the genetic relationship between the morphotypes by means of phylogenetic analyses using complete DNA sequences of the cytochrome b gene. The relationship between otolith weight and SL was significantly different between specimens 550 mm SL. Phylogenetic analysis demonstrated that the sequences from these scombropid specimens formed three clades: two corresponded to S. boops and S. gilberti, while the third did not correspond to any sequence recorded in databases, suggesting that these specimens are undescribed scombropid species. Almost all the specimens with SL n = 76) were identified as S. boops, and only nine as S. gilberti. On the other hand, almost all the specimens with SL > 550 mm (n = 41) fell in the unidentified group except for four specimens, whose sequences were identical to that of S. boops.

1 College of Bioresource Sciences, Nihon University, Fujisawa, Kanagawa 252-0880, Japan 2 Kanagawa Prefectural Museum of Natural History, Odawara, Kanagawa 250-0031, Japan 3 Yokohama Hakkeijima Sea Paradise, Yokohama, Kanagawa 236-0006, Japan 4 National Research Institute of Fisheries Science, Fisheries Research Agency, Yokohama, Kanagawa 236-8648, Japan 5 Japan Driftological Society, Hata, Kochi 789-1911, Japan

The Japanese gnomefishes Scombrops boops and S. gilberti closely resemble each other but their geographical distributions in the northwestern Pacific Ocean are markedly differ- ent. In order to understand the determinants of reproductive isolation, we examined habitat use in relation to the reproductive strategy employed by these species in the Izu-Islands region, where adults of both species coexist. We examined the species compositions of the gnomefishes based on mitochondrial sequence differences, and the maturation process of gonads using the gonado - somatic index (GSI) and by histological observation of the gonads. Genetic analysis showed that large individuals (>400 mm SL) of both species were present in the region, whereas all small gnomefish (

The complete mitochondrial genome of the Japanese gnomefish, Scombrops gilberti, was determined using a PCR-based method. The total length of mitochondrial DNA (mtDNA) is 16 518 bp, which includes 13 protein-coding genes, two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and one control region. The mitochondrial gene arrangement of S. gilberti was found to be identical to that of other scombropid and indeed, other teleosts as well. Maximum likelihood analysis revealed that Scombropidae forms a sister group to Pempheriformes.

The potential anticancer agent 1-(2-chlorophenyl-N-methylpropyl)-3-isoquinolinecarboxamide (PK11195), a translocator protein ligand (initially described as a ligand for the peripheral benzodiazepine receptor), induces apoptosis in some lines of human tumor cells. We investigated the effect of PK11195 in the choriocarcinoma cell line, BeWo. BeWo cells were treated with various concentrations of PK11195, and changes in cell growth, the cell cycle, apoptosis, and related parameters were examined. A WST-1 assay showed that BeWo cells were sensitive to the growth inhibitory effect of PK11195. In contrast, the nonsite selective ligand diazepam has a little effect on these cells. Cell cycle analysis indicated that exposure to PK11195 decreased the proportion of cells in the S phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by Annexin V staining of externalized phosphatidylserine, by the loss of mitochondrial transmembrane potential, and by antibodies directed against histones from fragmented DNA. This induction occurred in conjunction with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. These results suggest that PK11195 may serve as a therapeutic agent for the treatment of choriocarcinoma.

The morphological similarity between upstream fatminnow Rhynchocypris oxycephalus and downstream fatminnow Rhynchocypris lagowskii makes it difficult to discriminate accurately between these species in rivers where they coexist. For easy and precise identification of these two species, we developed a genetic discrimination method based on PCR-RFLP analysis for specimens from the Inohzawa River watershed in the Izu Peninsula, central Honshu, Japan. This genetic method was applied to the species identification of the fatminnows from two other watersheds, the Kano and Kawazu Rivers, flowing across the peninsula from north to south. We present the genetic evidence for the restricted distribution of R. oxycephalus and the ubiquitous distribution of R. lagowskii in the peninsula.

Bufalin is a traditional oriental medicines which induces apoptosis in some lines of human tumor cells. It constitutes the major digoxin-like immunoreactive component of Chan Su, obtained from the skin and parotid venom glands of toads. Bufalin is cardioactive C-24 steroids that exhibits a variety of biological activities, such as cardiotonic, anaesthetic, blood pressure stimulatory, respiratory and antineoplastic effects. In terms of its anti-tumor activity, bufalin has been demonstrated to inhibit the growth of tumors, such as endometrial and ovarian cancers. This commentary introduces biologic and therapeutic effects of bufalin in treating some cancers. The compound is able to mediate inhibition of cell growth, cell cycle arrest, apoptosis, and expression of genes related to the malignant phenotype in human cancer cells.

KN-93, a membrane-permeant calcium/calmodulin- dependent kinase-selective inhibitor, induces apoptosis in some lines of human tumor cells. We investigated the effect of KN-93 in the choriocarcinoma cell line, BeWo. BeWo cells were treated with various concentrations of KN-93, and changes in cell growth, the cell cycle, apoptosis, and related parameters were examined. A WST-1 assay showed that BeWo cells were sensitive to the growth inhibitory effect of KN-93. Cell cycle analysis indicated that exposure to KN-93 decreased the proportion of cells in the S phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by Annexin V staining of externalized phosphatidylserine, by the loss of mitochondrial transmembrane potential, and by antibodies directed against histones from fragmented DNA. This induction occurred in conjunction with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. These results suggest that KN-93 may serve as a therapeutic agent for the treatment of choriocarcinoma.

Scombrops gilberti is a member of the percoid family Scombropidae, which includes a single genus and three to four species worldwide. Little is known about the ecology of this species. Juvenile S. gilberti have been found in the waters off northern Japan (Iwate Prefecture), whereas adults are found in the waters around the southern counterpart (Izu Islands), approximately 700 km from the northern waters. In the present study, we captured immature S. gilberti (106–248 mm standard length, SL) in the northern waters by set net at 8–80 m depth, whereas larger individuals (150–328 mm SL) were captured by trawling at 150–500 m depth. By contrast, only adult S. gilberti (422–590 mm SL) were captured in the southern waters. The genetic composition of the adult population of S. gilberti from the southern waters and of the juvenile population from the northern waters was compared using the nucleotide sequence of the mitochondrial DNA (mtDNA) cytochrome b gene. No significant differences in genetic parameters such as fixation index, neutrality test or mismatch distribution analysis were found between these geographically distinct populations of S. gilberti, showing that these populations are genetically homogeneous.

Objective: Bufalin is a traditional Chinese medicine, and it induces apoptosis in some lines of human tumor cells. Methods: We investigated the effect of bufalin in the choriocarcinoma cell line, BeWo. BeWo cells were treated with various concentrations of bufalin, and changes in cell growth, the cell cycle, apoptosis, and related parameters were examined. Results: An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that BeWo cells were sensitive to the growth inhibitory effect of bufalin. Cell cycle analysis indicated that exposure to bufalin decreased the proportion of cells in the synthesis phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and by the loss of mitochondrial transmembrane potential. This induction occurred in conjunction with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. Conclusions: These results suggest that bufalin may serve as a therapeutic agent for the treatment of choriocarcinoma.

Primary small cell carcinoma of the endometrium is rare and has an extremely poor prognosis. This report describes two cases of small cell carcinoma of the endometrium diagnosed as stage III. Case 1 was diagnosed as stage IIIc. She underwent surgery and chemotherapy. For a locally recurrent tumor, she received radiotherapy. She has been well with no evidence of disease for 4 years. Case 2 was diagnosed as stage IIIa. She underwent surgery. The tumor recurred soon after the surgery, and she died 33 days after the surgery. In the literature, the median survival reported for patients with stage III and IV is only 5 months. Case 1 is the 4th case showing long-term survival with advanced-stage disease. The optimal treatment for this rare tumor has not been established. Considering its rarity and variability, it is difficult to establish an evidence-based therapeutic regimen.

background: Accumulating evidence suggests that various epigenetic aberrations play definite roles in the pathogenesis of endometriosis. We investigated the histone acetylation status in endometriosis and the application of the histone deacetylase inhibitors (HDACIs) for the treatment of endometriosis. methods: The levels of acetylated histones in the endometriotic cyst stromal cells (ECSCs) and normal endometrial stromal cells (NESCs) were evaluated. The effects of the HDACIs on cell proliferation, the cell cycle, apoptosis of ECSCs and NESCs, and the expression of genes related to these cellular events were investigated. The effects of HDACIs on histone acetylation in chromatin of the promoter region of the cell cycle regulatory genes in ECSCs were also investigated. results: The acetylated histone levels were significantly lower in ECSCs than in NESCs (P , 0.025). HDACIs inhibited cell proliferation and induced cell cycle arrest and apoptosis of ECSCs. The effects of HDACIs on NESCs were marginal or weak. These HDACIs induced an accumulation of acetylated histones in total cellular chromatin and in the promoter regions of the p16 INK4a , p21 Waf1/Cip1 , p27 Kip1 and cycle checkpoint kinase 2 genes in ECSCs. HDACIs induced the protein expression of these cell cycle regulators and suppressed the protein expression of Bcl-2 and Bcl-XL in ECSCs. conclusions: The present findings demonstrated that aberrant histone modifications are present in endometriosis and that HDACIs reactivated epigenetically silenced genes, resulting in the suppression of cell proliferation, induction of cell cycle arrest and apoptosis of ECSCs. HDACIs are therefore promising agents for the treatment of endometriosis.

Endometriosis, a common, benign, estrogen-dependent disease affecting 3-10% of women of reproductive age, is characterized by the ectopic growth of endometrial tissue that is found primarily in the peritoneum, ovaries and rectovaginal septum. Recently, endometriosis has been alternatively described as an immune disease, a genetic disease and a disease caused by exposure to environmental factors, in addition to its usual description as a hormonal disease. In addition, accumulating evidence suggests that various epigenetic aberrations play definite roles in the pathogenesis of endometriosis. Epigenetic alterations reported to date in endometriosis include the genomic DNA methylation of progesterone receptor-B, E-cadherin, homeobox A10, estrogen receptor-β, steroidogenic factor-1 and aromatase. Aberrant expression of DNA methyltransferases, which attach a methyl group to the 5-carbon position of cytosine bases in the CpG island of the promoter region and silence the corresponding gene expression, has also been demonstrated in endometriosis. This review summarizes the recent studies on the aberrant DNA methylation status and aberrant expression of DNA methyltransferases, which regulate DNA methylation, in endometriosis. We also discuss the recent information on the diagnostic and therapeutic implications of epigenetic alterations occurring in endometriosis.

Endometriosis, a disease affecting 3% to 10% of women of reproductive age, is characterized by the ectopic growth of endometrial tissue under the influence of estrogen. It is also becoming recognized as a condition in which ectopic endometrial cells exhibit abnormal proliferative and apoptotic regulation in response to appropriate stimuli. Apoptosis plays a critical role in maintaining tissue homeostasis and represents a normal function to eliminate excess or dysfunctional cells. Accumulated evidence suggests that, in healthy women, endometrial cells expelled during menstruation do not survive in ectopic locations because of programmed cell death, while decreased apoptosis may lead to the ectopic survival and implantation of these cells, resulting in the development of endometriosis. Both the inability of endometrial cells to transmit a "death" signal and the ability of endometrial cells to avoid cell death have been associated with increased expression of antiapoptotic factors and decreased expression of preapoptotic factors. Further investigations may elucidate the role of apoptosis-associated molecules in the pathogenesis of endometriosis. Medical treatment with apoptosis-inducing agents may be novel and promising therapeutic strategy for endometriosis.

A membrane-targeted, lipophilic ether lipid of synthetic phospholipid analog, erucylphosphocholine (ErPC), induces apoptosis in some lines of human tumor cells. We investigated the effect of ErPC in the choriocarcinoma cell line, BeWo. BeWo cells were treated with various concentrations of ErPC, and changes in cell growth, the cell cycle, apoptosis, and related parameters were examined. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that BeWo cells were sensitive to the growth inhibitory effect of ErPC. Cell cycle analysis indicated that exposure to ErPC decreased the proportion of cells in the S phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by Annexin V staining of externalized phosphatidylserine and by the loss of mitochondrial transmembrane potential. This induction occurred in conjunction with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. These results suggest that ErPC may serve as a therapeutic agent for the treatment of choriocarcinoma.

Because epigenetic alterations are believed to be involved in the repression of tumor suppressor genes and promotion of tumorigenesis in choriocarcinomas, novel compounds endowed with a histone deacetylase (HDAC) inhibitory activity are an attractive therapeutic approach. HDAC inhibitors (HDACIs) were able to mediate inhibition of cell growth, cell cycle arrest, apoptosis, and the expression of genes related to the malignant phenotype in choriocarcinoma cell lines. In this review, we discuss the biologic and therapeutic effects of HDACIs in treating choriocarcinoma, with a special focus on preclinical studies.

Some macrophytes are transported to the deep-sea bottom and are utilized by heterotrophs in the deep-sea as a food source. We inferred the transport route of macrophytes toward the deep-sea based on similarity in the species compositions of macrophyte pieces collected from the dysphotic bottom off the Izu Peninsula and the drifting macroalgae reported for the study area. We also examined whether or not the macrophytes are buried in the sediment, based on stable isotope distributions of organisms. Macrophytes collected by dredging at a depth of 100–300 m included 93 species, whereas 43 species were found by trawling at depths from 200 to 400 m. Only 15 of 76 dredged species (19.7%) that were identified to the species level were identical to the drifting macroalgal species reported for this area, whereas 15 of the 29 trawled species (51.7%) that were identified to the species level were identical to the reported drifting species. It was thus inferred that macrophytes were mainly transported through sliding along the sea bottom for the macrophytes collected by dredging and through sinking from the surface water for the macrophytes collected by trawling. The δ13C of sedimentary organic matter (SOM) from the 200–300 m zone was similar to the δ13C distribution of particulate organic matter in the surface water reported for the study area. The SOM in the zone likely originated from almost exclusively phytoplankton. In contrast, the 13C of SOM was significantly more enriched in shallow areas ≤100 m deep. We infer that not only phytoplankton but also macrophytes could supply organic matter to heterotrophs on the shallow bottom.

Two scombropid fishes, Scombrops boops and Scombrops gilberti, are closely related and commercially important species in Japan. These species are often confused in commercial markets because of their morphological similarity. In this study, scombropid specimens collected from various Japanese coastal waters were subjected to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and phylogenetic analysis of the 16S rRNA gene in mitochondrial DNA. These analyses showed that all the scombropid specimens collected from localities in the Sea of Japan were identified as S. boops, whereas those from the Pacific Ocean included two species, S. boops and S. gilberti. Almost all juvenile (200 mm standard body length, S(L)) S. gilberti originated from the Pacific coastal waters of the northern Japan, whereas adults (400 mm S(L)) were found only in deep water off the Izu Peninsula to the Izu Islands. This suggests that S. gilberti might migrate extensively during its life cycle. In addition, differences in the number of specimens and the distribution between the two species suggest that S. gilberti is less abundant than S. boops in Japanese waters.

The auditory pressure thresholds of red sea bream were examined using cardiac response in the field by placing fish subjects far from the sound source to prevent particle motion. Pressure and particle motion thresholds were also obtained using the auditory brainstem response (ABR) technique. The thresholds at 100 and 200 Hz were significantly higher when measured using the cardiac response in the far field than those obtained in previously conducted experiments in experimental tub. However, thresholds obtained using ABR from 200 to 500 Hz were not remarkably lower, although significantly different (0.01

To investigate the efficacy of wide local excision as a surgical treatment for early-stage vulvar melanomas.Wide local excision with or without lymph node dissection was performed in three patients with stage I vulvar melanoma (American Joint Committee on Cancer classification, 1992).All three patients were successfully treated by wide local excision. There was no evidence of recurrence at long-term follow-up in any of the patients.Wide local excision with adequate tumor-free margins should be considered the treatment of choice for early-stage vulvar melanomas.

Objectives. A membrane-targeted, lipophilic ether lipid of synthetic phospholipid analog, erucylphosphocholine (ErPC) induces apoptosis in some lines of human tumor cells. We investigated the effect of ErPC on three endometrial cancer cell lines, two ovarian cancer cell lines, and normal human endometrial epithelial cells. Methods. Endometrial and ovarian cancer cells were treated with various concentrations of ErPC, and its effect on cell growth, cell cycle, apoptosis, and related measurements was investigated. Results. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that all endometrial and ovarian cancer cell lines were sensitive to the growth-inhibitory effect of ErPC, although normal endometrial epithelial cells were viable after treatment with the same doses of ErPC that induced growth inhibition in endometrial and ovarian cancer cells. Cell cycle analysis indicated that their exposure to ErPC decreased the proportion of cells in the S-phase and increased the proportion in the G2/M phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and loss of the transmembrane potential of mitochondria. This induction occurred in concert with altered expression of genes related to cell growth, malignant phenotype, and apoptosis. Conclusions. These results suggest that the anticancer activity of ErPC may occur with higher sensitivity of cancer cells compared with normal healthy cells, when using low concentration, rising hopes that ErPC may become a useful adjuvant therapy for endometrial and ovarian cancers.

Gnomefish Scombrops boops and Scombrops gilberti are commercially important fishes in Japan, but these species are often confused in the markets because of their morphological similarity. To identify these two species, we performed nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis on 16S ribosomal RNA (rRNA) gene and the control region in mitochondrial DNA. Five and 12 nucleotide substitutions were observed between species in the 777-bp 16S rRNA gene and 471-bp control region, respectively. Diagnostic restriction sites for discriminating between S. boops and S. gilberti were found in the 16S rRNA gene, but not in the control region. Polymerase chain reaction (PCR)-RFLP analysis using two enzymes, EcoNI and Mval, clearly discriminated between S. boops and S. gilberti identified by meristic characters. The PCR-RFLP analysis identified most of the 168 Scombrops young caught in the coastal waters of the Izu and Miura peninsulas as S. boops, suggesting that S. gilberti juveniles are rare in this area.

Objectives Beta-hydroxyisovalerylshikonin (beta-HIVS), a compound isolated from the traditional oriental medicinal herb Lithospermum radix, is an ATP non-competitive inhibitor of protein-tyrosine kinases, such as v-Src and EGFR, and it induces apoptosis in some lines of human tumor cells. We investigated the effect of beta-HIVS on three endometrial cancer cell lines, two ovarian cancer cell lines, and normal human endometrial epithelial cells. Methods Endometrial and ovarian cancer cells were treated with various concentrations of beta-HIVS, and its effect on cell growth, cell cycle, apoptosis, and related measurements was investigated. Results The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that all endometrial and ovarian cancer cell lines were sensitive to the growth-inhibitory effect of beta-HIVS, although normal endometrial epithelial cells were viable after treatment with the same doses of beta-HIVS that induced growth inhibition in endometrial and ovarian cancer cells. Cell-cycle analysis indicated that their exposure to beta-HIVS decreased the proportion of cells in the S phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and loss of the transmembrane potential of mitochondria. This induction occurred in concert with altered expression of genes related to cell growth, malignant phenotype, and apoptosis. Conclusions These results suggest that the anticancer activity of beta-HIVS may occur with higher sensitivity of cancer cells compared with normal healthy cells, when using low concentration, rising hopes that beta-HIVS may become a useful adjuvant therapy for endometrial and ovarian cancers.

Tamoxifen is widely used for endocrine treatment and breast cancer prevention. It acts as both an estrogen antagonist in breast tissue and an estrogen agonist in the female lower genital tract. Tamoxifen causes severe gynecologic side effects, such as endometrial cancer. This review focuses on the effects of prolonged tamoxifen treatment on the human female genital tract and considers its tumorigenicity in the gynecologic organs through clinical data analysis. Tamoxifen is associated with an increased incidence of benign endometrial lesions such as polyps and hyperplasia and a two- to four-fold increased risk of endometrial cancer in postmenopausal patients. Moreover, the incidence of functional ovarian cysts is significantly high in premenopausal tamoxifen users. To prevent tamoxifen from having severe side effects in gynecologic organs, frequent gynecological examination should be performed for both premenopausal and postmenopausal patients with breast cancer who are treated with this drug.

Because epigenetic alterations are believed to be involved in the repression of tumor suppressor genes and promotion of tumorigenesis in endometrial cancers and ovarian cancers, novel compounds endowed with a histone deacetylase (HDAC) inhibitory activity are an attractive therapeutic approach. Clonogenic assay in soft agar and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays showed that many endometrial and ovarian cancer cell lines were sensitive to the growth inhibitory effect of HDAC inhibitors (HDACIs), although normal endometrial epithelial cells were viable after the treatment with the same doses of HDACIs that induced growth inhibition of endometrial and ovarian cancer cells. Cell cycle analysis indicated that their exposure to HDACIs decreased the proportion of cells in the S-phase and increased the proportion in the G0/G1 phases and/or G2/M phases of the cell cycle. Induction of apoptosis was confirmed by TUNEL assay, annexin V staining of externalized phosphatidylserine, and loss of the transmembrane potential of mitochondria. This induction occurred in concert with altered expression of genes related to cell growth, malignant phenotype, and apoptosis. In nude mice experiments, valproic acid significantly inhibited human endometrial and ovarian tumor growth without toxic side-effects. Although there are few clinical trials on these cancers, some clinical trials showed that HDACIs in well tolerated doses have significant antitumoral activities in another cancers. These results raise the possibility that HDACIs may prove particularly effective in the treatment of endometrial cancers and ovarian cancers.

It is important to clarify trophic dynamics in marine ecosystems for management of the fishing ground. Organic carbon sources and trophic position of pelagic fishes in the coastal waters of the south-eastern Izu Peninsula, Japan, were examined on the basis of carbon and nitrogen stable isotope distributions. The δ13C of the fishes was mostly distributed from −19 to −16‰ for nektonic fishes (13 species of adults and immatures) and planktonic fishes (10 species of larvae and juveniles), close to the δ13C values of particulate organic matter and planktonic decapods. These δ13C signatures for the inhabitants of the water column were in contrast with the high δ13C values (mainly −16 to −13‰) for demersal fishes of Scorpaeniformes and benthic polychaetes collected in the surf zone. These results indicate that nektonic and planktonic fishes depend on phytoplankton for carbon supply. The δ15N signatures suggest that the trophic position ranged 3.1–4.5 for the nektonic fishes and 2.9–3.7 for the planktonic fishes, premised on trophic level 3 for larval Japanese anchovy Engraulis japonicus. Thus, planktivorous fishes should be mainly assigned to trophic levels 3 and 4 in this area.

Girella punctata and Girella leonina are sympatric sister species showing extensive distributional overlap in shallow rocky reefs in the Pacific Ocean south of the Japanese Islands. Differences between the two species in external morphological characters, such as number of pored lateral line scales, colour of opercular flap and shape of caudal fin, are congruent with genetic divergence. Nucleotide identity between the two species in the 3.3 kbp region of partial mitochondrial DNA containing the D-loop region, in 12S and 16S ribosomal RNA (rRNA) and transfer RNA genes is 95%. To estimate divergence time, Bayesian analysis was conducted using a dataset comprising concatenated nucleotide sequences from the two rRNA genes of three girellid and nine other fish species. Using the Elopomorpha – Clupeocephala split (265 million years ago (mya)) as a calibration point, divergence between G. punctata and G. leonina is estimated as having occurred 6.0±1.4 mya. Speciation is suggested to have been caused by geographical isolation associated with formation of the Japanese Islands, which resulted in disjunction of Girella habitat.

The contribution of benthic microalgal production has been compared both within and outside a coastal eelgrass ( Zostera marina L.) meadow. Carbon and nitrogen stable isotope ratios of suspended particulate organic matter (POM), epiphytic and epilithic organic matter (EOM), leaves of Z. marina (inside the meadow only) and two secondary consumer species (small crustaceans and fish) were measured inside and outside a meadow in Mitsukuchi Bay, Northwest Seto Inland Sea, Japan. Inside the meadow, primary producers (epiphyton) and consumers showed higher δ 13 C signatures than outside. Primary and secondary consumers inside the meadow were mainly dependent on epiphyton on the leaves of Z. marina , while consumer species outside the meadow were basically dependent on epilithon.

Background Most of the current medical treatments for endometriosis aim to down-regulate the estrogen activity. However, a high recurrence rate after medical treatments has been the most significant problem. Beta-hydroxyisovalerylshikonin (beta-HIVS) is an ATP non-competitive inhibitor of protein-tyrosine kinases and is considered an apoptosis-inducing agent. The aim of this study is to evaluate the effects of beta-HIVS on the proliferation, cell cycle and apoptosis of endometriotic stromal cells. Methods We investigated the effects of beta-HIVS on cultured ovarian endometriotic cyst stromal cells (ECSC) by a modified methylthiazoletetrazolium (MTT) assay, a 5-bromo-2'-deoxyuridine (BrdU) incorporation assay and internucleosomal DNA fragmentation assays. The effect of beta-HIVS on the cell cycle of ECSC was determined by flow cytometry. The expression of apoptosis-related molecules was examined in ECSC using western blot analysis. Results Beta-HIVS significantly inhibited the proliferation and DNA synthesis of ECSC and induced apoptosis and G0/G1 phase cell-cycle arrest of these cells. Down-regulation of the B-cell lymphoma/leukaemia-2 (Bcl-2) expression with the activation of caspase-3, caspase-8 and caspase-9 was observed in ECSC after beta-HIVS treatment. Conclusions These results suggest that beta-HIVS induces apoptosis of ECSC by suppressing anti-apoptotic proteins. Although our present findings are preliminary, beta-HIVS could potentially be a therapeutic agent for the treatment of endometriosis.

Stable carbon and nitrogen isotope ratios were determined in the baleen plates of 17 common minke whales Balae noptera acutorostrata from the north-western Pacific Ocean off Japan, as well as prey species (krill Euphausia pacifica, Japanese anchovy Engraulis japonicus and Pacific saury Cololabis saira) collected in the stomach contents, to investigate the trophic relationship between the minke whales and their prey. A few δ15N-depleted peaks occurred along the length of baleen plates for 10 males irrespective of stomach content (anchovies and sauries). Similar δ15N-depleted peaks were also found for one female and two immature individuals. It was likely that these δ15N-depleted peaks formed in early summer. The stable nitrogen isotope ratio (δ15N) values in Pacific saury (9.3±1.4‰) did not differ significantly from that in Japanese anchovy (8.8±0.9‰). In contrast. δ15N in krill (7.2±0.5‰ in July and 8.0±0.2‰ in September) were significantly lower than in the Pacific saury. Thus, these peaks may reflect the dietary change from krill to fishes in the feeding migration of the whales. Growth rate of the baleen plate was estimated to be 129 mm/y, and it appeared that a dietary record of about 1.4 years remained in the baleen plate. For two immature whales, the maximum value of δ15N occurred at the tip of baleen. This δ15N enrichment may possibly be useful for discriminating weanlings and older whales.

Objectives We investigated the effect of a novel synthesized histone deacetylase inhibitor (HDACI), CBHA, on three endometrial cancer cell lines, two ovarian cancer cell lines, and normal human endometrial epithelial cells. Methods Endometrial and ovarian cancer cells were treated with various concentrations of CBHA, and its effect on cell growth, cell cycle, apoptosis, and related measurements was investigated. Results The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that all endometrial and ovarian cancer cell lines were sensitive to the growth-inhibitory effect of CBHA, although normal endometrial epithelial cells were viable after treatment with the same doses of CBHA that induced growth inhibition in endometrial and ovarian cancer cells. Cell cycle analysis indicated that their exposure to CBHA decreased the proportion of cells in the S-phase and increased the proportion in the G(0)/G(1) phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and loss of the transmembrane potential of mitochondria. This induction occurred in concert with altered expression of genes related to cell growth, malignant phenotype, and apoptosis. Furthermore, CBHA treatment of these cell lines increased acetylation of H3 and H4 histone tails. Conclusions These results raise the possibility that CBHA may prove particularly effective in the treatment of endometrial and ovarian cancers.

BACKGROUND Human epidermal growth factor receptor 2 (HER2) is overexpressed in 25–30% of ovarian carcinoma cases and a correlation between increased HER2 expression and decreased survival has been demonstrated. HER2 is a ligand-less member of the HER family that functions as the preferred coreceptor for epidermal growth factor receptor (EGFR), HER3, and HER4. METHODS An approach was developed to target HER2's role as a coreceptor using a monoclonal antibody, 2C4, which sterically hinders HER2's recruitment into a functional HER complex. RESULTS HER2 was robustly expressed in all eight ovarian carcinoma cell lines; expression of EGFR and HER3 was variable. Even though four of the eight cell lines responded to EGF, 2C4 antibody moderately inhibited in vitro proliferation of only two cell lines (OVCA433 and SK-OV-3). Furthermore, ligand-stimulated p-MAPK expression was inhibited by 2C4 only in these two cell lines after exposure to EGF. Immunoprecipitation and eTag analysis revealed that OVCA433 expressed heterodimers of EGFR/HER2, and these heterodimers were disrupted after treatment with 2C4, whereas OVCA432 cells did not have these heterodimers. In murine xenograft experiments, the in vivo growth of OVCA433, but not of OVCA432, ovarian carcinoma cells was significantly inhibited by 2C4 treatment of the mice. CONCLUSION 2C4 is able to disrupt the HER signaling pathway and inhibit the in vitro and in vivo growth of ovarian carcinoma cell lines. The response appears limited to lines in which HER2 heterodimers were able to transduce proliferative signals. Our findings suggest a strong rationale to conduct clinical trials of 2C4 in a subset of patients with ovarian tumors. Cancer 2005. © 2005 American Cancer Society.