The purposes of this study were to establish the reference values of element concentrations and to estimate the food safety and the nutrient values of essential elements in the aquaculture fish products in Taiwan. 31 species of fishes, 4 species of crustaceans and 3 species of molluscs were collected from 11 aquaculture areas in Taiwan from September to November in 2004 to 2006. Concentrations of Ag, As, Cd, Co, Cu, Fe, Hg, Mn, Se, Sr and Zn in the muscles were analyzed. The results showed that the concentrations of those elements had significant difference among teleosteans, crustaceans and molluscs, excepted Hg and Se. Essential element concentrations in Cu, Sr and Zn were higher in invertebrates (shellfishes) than in vertebrates (teleosteans) but Co, Fe and Mn were higher in molluscs than others. All concentrations of non-essential elements excepted Hg were higher in molluscs. Significant species-differences were also found in our results, but not in the concentrations of Ag and Co in teleosteans. Among teleosteans, Trachinotus blochii (As and Se), Anguilla japonica (Cd and Zn), Lates calcarifer (Cu), Carassius auratus auratus (Fe), Epinephelus coioides (Hg), Rachycentron canadum (Hg) and Chanos chanos (Mn and Sr) contained the highest concentrations, showed in parenthesis. Whereas among shellfishes, Haliotis diversicolor (Ag), Meretrix lusoria (As, Cd, Co, Fe, Mn, Se and Sr), Corbicula fluminea (Co and Se), Scylla serrata (Hg and Zn) revealed the highest concentrations of those elements. The results were seminar to previous literatures reported for uncontaminated aquatic environment area worldwide, so they could be the reference values of element concentrations in the aquaculture fish products in Taiwan. The element concentrations in this study were all below the worldwide food safety standards of inorganic As = 1.0, Cd (for fish and crustaceans) = 0.2, Cd (for mollusks) = 2.0, Cu = 10, total Hg = 0.5, Se = 1.0 and Zn = 50 £gg / g wet wt. Based on the normal dietary habit of Taiwanese by consume 400g fishes and 200g shellfishes every week, the element concentrations of weekly intake were bellowed the standards of inorganic As = 15, Cd = 7 and organic Hg = 1.6 £gg / kg of body / week by PTWI, and could obtain more than 100% to Co and 58% to Se of RDA standards at the same time.

The objects of this study were to investigate the differences of total mercury(THg), organic mercury(OHg) concentrations in the muscles of albacore and bigeye tuna from the Pacific Ocean, and the THg and OHg concentration in the livers , and the trace elements (Cu, Fe, Mn and Zn) in the muscles and livers of albacore, in order to evaluate the bioaccumulative status of the oceanic migratory fishes. Meanwhile, The relationships between mercury concentration and size (length and weight) and age were established, and the differences of mercury concentrations in the tuna between species and geography were examined. And that the safety consumption level was also discussed. One hundred eighty-five albacore and 134 bigeye tuna were sampled from the period October 2001 to April 2006 and from September 2005 to November 2006, respectively, in the Western and Central Pacific Ocean. The results showed that the average THg (mg/kg flesh wt.) and OHg (mg/kg flesh wt.) of muscles were 0.435¡Ó0.145 and 0.279¡Ó0.087, respectively, for albacore, and 0.935¡Ó0.655 and 0.544¡Ó0.396, respectively, for bigeye tuna. The THg and OHg of livers of albacore were 0.426¡Ó0.363 and 0.193¡Ó0.121, respectively. The average OHg percentages of the THg in albacore and bigeye tuna were 67¡Ó19 % and 62¡Ó20 %, respectively. Except there was no significant correlation between OHg and age in the liver of albacore, the THg and OHg in the muscles, and THg in the livers of albacore, and THg in the muscles of bigeye tuna showed a positive correlation with size and age. The THg and OHg levels in the muscles of albacore were lower than those of bigeye tuna, and the accumulative rates of THg and OHg in albacore were lower than those in bigeye tuna. No significant different of muscles-THg and -OHg concentrations of albacore from the four samplind areas. However, the THg and OHg levels in muscle of bigeye tuna showed that north group higher than those in equator. The concentrations of THg were similar in muscle and liver of albacore, but those OHg, Cu, Fe, Mn and Zn concentrations in liver were significantly higher than those in muscle. A detoxificated mechanism may be commenced when the concentration of THg in liver of albacore exceed 0.8 mg/kg, to maintain the muscle concentration of THg at a level about 0.7 mg/kg. The OHg concentrations of all albacore and 88% of bigeye tuna were below WHO food safety standard for migratory fishes(1.0 mg/kg). The consumption amount of albacore and bigeye tuna were no more than 300 and 150g, respectively, will not exceesed the PTWI(provisional tolerable weekly intake) set by WHO, that will not cause any health threat for consumer.

The parmacological effects of guinea pig vasoactive intestinal peptide (VIP) were studied in isolated perfused guinea pig hearts. Bolus injections of VIP produced a dose-dependent tachycardia that was not affected by atenolol. A decrease in amplitude of ventricular contractions occurred in response to all doses of VIP. This response was preceded by a small increase in amplitude in 3 of 6 hearts at the highest dose. VIP produced a decrease in perfusion pressure which was prominent after coronary tone was elevated with [Arg8]-vasopressin. The present findings support speculation that VIP may have a role in the regulation of heart rate and coronary blood flow.

京都大学 / 0048 / 新制・論文博士 / 博士(農学) / 乙第12834号 / 論農博第2798号 / 新制||農||1026(附属図書館) / 学位論文||H26||N4857(農学部図書室) / 31372 / (主査)教授 栗原 達夫, 教授 植田 和光, 教授 平竹 潤 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

The objectives of this study were to investigate the difference and interaction of mercury (Hg) and selenium (Se) among fish species and tissues. In addition, by comparing with the detected concentrations in other regions, it would be possible to evaluate the pollution status. In the meanwhile, this study analyzed the edibility of the fish, the interaction between Hg and Se as well as their impacts on fish growth. Ten species of fish, including tenpounder (Elops machnata), fourlined terapon (Pelates quadrilineatus), whipfin silverbiddy (Gerres filamentosus), common ponyfish (Leiognathus equulus), Hamilton's thryssa (Thryssa hamiltonii), tilapia (Orechromis niloticus), western Pacific gizzard shad (Nematalosa come), large-scaled mullet (Liza macrolepis), flathead mullet (Mugil cephalus) and milkfish (Chanos chanos), and 20 sediment samples were collected from September to October of 2003 at the reservoir adjacent to a chlor-alkali plant, which had been abandoned for 22 years in Tainan City. Total mercury (THg) and organic mercury (OHg) concentrations were determined in fish muscles, livers, kidneys, gonads, gills and sediment samples. Besides, Se concentrations were also analyzed in fish muscles and livers. Concentrations (mg/kg dry wt.) of THg and OHg in sediment were 43.2¡Ó23.4(mean¡Ósd) and 0.013¡Ó0.008, respectively. THg, OHg and Se concentrations (mg/kg wet wt.) in fish muscles were 0.432¡Ó0.360, 0.305¡Ó0.206 and 0.126¡Ó0.030, similar to the concentrations in other chlor-alkali polluted regions. With the diet habit of Taiwanese, the Hg concentrations of muscles in the seven fish species exceeded the limit of Hg which was allowed to be consumed by humans. THg and OHg concentrations in the fish muscles showed significant species difference. Obviously, the concentrations in carnivorous fishes were higher than those in omnivorous and herbivorous fishes. The OHg concentrations of fish livers and kidneys were highest in carnivorous fishes, but the THg concentrations were higher in omnivorous fishes than in that of carnivorous and herbivorous fishes. Besides, Se concentrations in fish muscles and livers also showed significant species difference, but did not show relation with the feeding habit of fishes. The Hg levels among fish tissues were highest in livers or kidneys, followed by gonads and muscles, lowest in gills. Generally speaking, Se levels were higher in livers than in muscles. The liver THg concentrations of tilapia reached 10 mg/kg wet wt. Their muscle THg concentrations decreased suddenly and maintained at a level about 0.4 mg/kg wet wt. Meanwhile, the liver inorganic Hg and Se concentrations increased with fish weight, while the liver OHg concentrations did not show such a trend but stay at a low level. High Hg concentrations in the fish tissues resulted in adverse effects on fish health. High Hg concentrations in muscles and livers caused heptasomatic index to decrease in western Pacific gizzard shad and milkfish, and condition factor declined in tilipia. Moreover, gonasomatic index remained low while the Hg concentrations of muscles, livers and gonads reached a threshold, which suggested that the growth of those fishes was likely inhibited in Hg polluted environment.

Preclinical testing is an important part of cancer drug development. The aim of this thesis was to establish and evaluate preclinical in vitro methods useful in the development of new anticancer drugs. In paper I, the development of non-clonogenic assays (FMCA-GM) using CD34+ stem cells for assessment of haematological toxicity was described. A high correlation was seen when comparing the 50% inhibitory concentrations (IC50) from FMCA-GM with the IC50 from the established clonogenic assay (CFU-GM). In paper II, FMCA-GM was complemented with additional cell models, establishing a normal cell panel. In vitro toxicity towards the five normal cell types was compared with known clinical adverse event profiles. The normal cell panel roughly reflected the tissue specific toxicities but was most useful in the prediction of therapeutic index. In paper III the use of peripheral blood lymphocytes from human, dog, rat and mouse to detect species differences in cellular drug sensitivity was described. Good agreement between our method and the established CFU-GM assay was observed. In paper II the benefit of using primary tumour cells from patients to predict cancer diagnosis-specific activity was studied. The in vitro activity of fourteen anticancer drugs was tested in tumour samples of both haematological and solid tumour origin. In general, clinical activity was well reflected. In paper IV, the efficacy and toxicity models were applied for experimental follow-up of a novel inhibitor of the ubiquitin-proteasome system, CB3 (Phosphoric acid, 2,3-dihydro-1,1-dioxido-3-thienyl diphenyl ester). In the preliminary characterization of CB3, antitumour activity and a favourable toxicity profile were displayed, although the exact mechanism of action remains to be elucidated. CB3 will therefore be further investigated. In conclusion, the work presented here contributes to different parts of the preclinical drug development and the methods may aid in the characterization of anticancer compounds

Depuis plusieurs années, un nombre conséquent d’études décrivent une augmentation de l’incidence de pathologies liées à la fonction de reproduction masculine. Ces anomalies ont été regroupées sous le terme de « syndrome de dysgénésie testiculaire ». Ce syndrome aurait pour origine les effets délétères de polluants environnementaux sur le développement du testicule en période fœtale. Parmi ces polluants environnementaux, les phtalates et le bisphénol A (BPA) sont les plastifiants les plus produits et les plus répandus dans les objets de consommation courante. De nombreuses études leur sont consacrées et ont permis de les classer au rang de perturbateurs endocriniens en mettant notamment en cause leurs effets reprotoxiques. Mon travail de thèse est une étude des effets de ces deux perturbateurs endocriniens sur le développement du testicule fœtal.Nous avons réalisé une première étude concernant les effets du BPA sur le développement du testicule fœtal. Grâce au modèle de culture organotypique, nous avons développé notre étude dans trois espèces : le rat, la souris et l’Homme. Nous démontrons que le BPA diminue la sécrétion de testostérone dans le testicule fœtal humain à partir d’une concentration de 10-8M, alors que chez le rat et la souris, la sécrétion de testostérone n’est affectée qu’à partir de 10-5M de BPA. Nous avons également démontré une diminution de l’expression du gène de l’Insl-3, dans ces mêmes conditions. Ceci nous a permis de mettre en évidence une différence de sensibilité entre les espèces. Pour tenter de comprendre les mécanismes par lesquels le BPA exerce son effet toxique, nous avons comparé ses effets à ceux du DES, autre perturbateur endocrinien œstrogénomimétique. Contrairement au BPA, le DES diminue la sécrétion de testostérone fœtale chez les rongeurs, et non chez l’Homme. Ce résultat suggère l’implication de deux voies de signalisation différentes pour ces deux xéno-œstrogènes. Cette hypothèse est d’ailleurs renforcée par l’étude que nous avons SourceMécanismes d’action des perturbateurs endocriniens bisphénol A et phtalates sur le développement du testicule fœtal / par Thierry N’Tumba-Byn ; sous la direction de Virginie Rouiller-Fabre, Université Paris Sud, 2013 [Thèse de Biologie de la Reproduction et du Développement]réalisée sur des souris invalidées pour le récepteur des œstrogènes ERα, dans lesquelles l’effet anti-androgénique du BPA persiste, contrairement à celui du DES.Parallèlement, nous avons recherché les mécanismes d’action des phtalates et de leur métabolite actif le plus répandu, le MEHP (mono-2-éthyl-hexyl phtalate). Dans la continuité de plusieurs travaux réalisés dans notre laboratoire sur les effets du MEHP, nous avons tenté de comprendre les mécanismes par lesquels le MEHP induit un effet pro-apoptotique dans les cellules germinales mâles. Nous avons mis en évidence une augmentation de l’expression du gène Stra8 dans les cellules germinales traitées au MEHP. Ce résultat nous suggère que le MEHP pourrait induire une différenciation erronée des cellules germinales mâles. De plus, nous avons recherché les récepteurs et la voie de signalisation activée par le MEHP. Nous observons que les agonistes des récepteurs PPARα et de PPARγ entrainent dans les cellules germinales les mêmes phénotypes que le MEHP, à savoir une augmentation du taux d’apoptose et de l’expression du gène Stra8. / For several years, an increase in the incidence of pathologies connected to the male reproductive functions has been described in numerous studies. These anomalies are classified under the term “testicular dysgenesis syndrome”. This syndrome might find its origins in the deleterious effects of environmental pollutants on the testis development in fetal period. Among theses environmental pollutants, phthalates and bisphenol A (BPA) are the most produced plasticizers found in products of common use. Many studies were performed in order to determine their effects, and allowed to classify them as endocrine disruptors because of their reprotoxic effects. My thesis work is a study of the effects of these two endocrine disruptors on the fetal testis development.Our first study focuses on the effects of BPA on the fetal testis development. Using the organotypic culture model, we developed our study in three species: rat, mouse and human. We demonstrated that BPA decreases the testosterone secretion in the human fetal testis from a 10-8M concentration, while in rat and mouse the testosterone secretion is only affected by 10-5M BPA. We also demonstrated a decreased Insl-3 gene expression, in the same conditions. These results allowed us to evidence a difference of sensibility between species. To understand the mechanisms involved in the BPA toxic effect, we compared it with the effect of DES, another endocrine disruptor with estrogenic activity. Unlike BPA, DES decreases the fetal testosterone secretion in rodents and not in human. This result suggests the involvement of two different signalisation pathways for these two xenoestrogens. This hypothesis is reinforced by the study that we performed in mice invalidated for the estrogen receptor ERα. In those mice, the anti-androgenic effect of BPA is maintained, unlike DES effect.In parallel, we investigated the mechanisms of action of phtalates and particularly of their most prevalent active metabolite, the MEHP (mono-2-ethyl-hexyl phthalate). Following previous studies performed in our laboratory concerning the effects of MEHP, we intended to understand the mechanisms by which MEHP induces the apoptosis in male germ cells. We evidenced an increase in Stra8 gene expression in MEHP treated germ cells. This result suggests that MEHP might induce a wrong differentiation in male germ cells. Furthermore, we investigated the receptors and the signalisation pathway activated by MEHP. We observe that PPARα and PPARγ receptors agonists induce the same phenotypes as MEHP, namely an increase in the apoptosis and in Stra8 gene expression in germ cells.