Genetic diversity and structure of urban populations of Pieris butterflies assessed using amplified fragment length polymorphism

Conservation programs in urban ecosystems need to determine the genetic background in populations of urban dwellers. We examined the genetic diversity and structure of Pieris rapae and P. melete using AFLP markers, and compared them between species and between urban and rural environments. As a result: (i). in both species, there was no reduction in genetic diversity within urban populations by direct comparison of diversity measurements, although the analysis of molecular variance suggested significant reductions in the variance within seasonal subpopulations in urban populations; (ii). P. rapae retained greater genetic diversity within species and populations; (iii). populations of both species showed significant genetic differentiation, and P. melete was more strongly subdivided; (iv). in both species, geographically close populations did not cluster with one another in the upgma analysis; (v). there was no genetic isolation due to geographical distance in either species; (vi). the genetic composition of seasonal subpopulations differed in urban populations of both species, and the genetic distances among subpopulations were correlated with seasonal differences in P. rapae and with temporal differences in P. melete. These results indicate that the genetic diversity in urban populations of both species was reduced at times, but was maintained by dispersal from genetically differentiated populations. Differences in the ability and mode of dispersal in the two species may be reflected in the degree of population subdivision and patterns of seasonal change in the genetic composition.     

Over-expression of a barley aquaporin increased the shoot/root ratio and raised salt sensitivity in transgenic rice plants

Barley HvPIP2;1 is a plasma membrane aquaporin and its expression was down-regulated after salt stress in barley [Katsuhara et al. (2002) Plant Cell Physiol. 43: 885]. We produced and analyzed transgenic rice plants over-expressing barley HvPIP2;1 in the present study. Over-expression of HvPIP2;1 increased (1) radial hydraulic conductivity of roots (Lp(r)) to 140%, and (2) the mass ratio of shoot to root up to 150%. In these transgenic rice plants under salt stress of 100 mM NaCl, growth reduction was greater than in non-transgenic plants. A decrease in shoot water content (from 79% to 61%) and reduction of root mass or shoot mass (both less than 40% of non-stressed plants) were observed in transgenic plants under salt stress for 2 weeks. These results indicated that over-expression of HvPIP2;1 makes rice plants sensitive to 100 mM NaCl. The possible involvement of aquaporins in salt tolerance is discussed.     

Proportion of mated females, female mating experience, and sex ratio of the osmund sawfly, Strongylogaster osmundae (Hymenoptera, Tenthredinidae)

The proportion of mated females (M _f) of the osmund sawfly, Strongylogaster osmundae, and the sex ratio of the eggs they deposited (r, proportion of males) were estimated in the wild by collecting egg masses. The proportion of mated females at oviposition varied from 0 to 1.0. M _f was high (often 1.0) among the females that emerged after hibernation, and lower in the subsequent generations. Mated females of the hibernated generation deposited equal numbers of eggs of both sexes. Mated females of the first and subsequent generations produced more female than male eggs. These results qualitatively agreed with the prediction provided by an evolutionarily stable strategy (ESS) model (if M _f < 1 then r < 0.5). However, the quantitative prediction provided by the model [M _f (1 − r) = 0.5] was not always observed in the wild, especially where the population density and M _f were high. The value of r was often lower than the predicted one. The following simple hypothesis was tested by experimentation: “Females that encounter males frequently estimate the proportion of mated females to be high and deposit eggs with a 1:1 sex ratio.” However, results did not support this hypothesis. Females that copulated soon after emergence and were courted by males two or more times did not show a higher offspring sex ratio than those which mated 1 or 2 days after emergence and experienced no other sexual encounter. Another mechanism for determination of r is suggested, and the reason why the population sex ratio of sawflies is often female-biased (r < 0.5) is discussed.     

Induction of Browning of Male Flowers of Cryptomeria japonica by Treatment with Fatty Acids: Mechanism and the Role of trans-2-Hexenal

The effect of treatment with fatty acids on the browning ofmale flowers of Cryptomeria japonica and its mechanism werestudied with emphasis on the role of trans-2-hexenal. The effectof treatment with fatty acids on the browning of male flowerswas the highest when fatty acids were applied at the end ofAugust, decreasing month by month thereafter in parallel withthe seasonal changes in lipoxygenase activity. trans-2-Hexenal,which is formed from linolenic acid in a reaction catalyzedby lipoxygenase (EC 1.13.1.13 [EC] ), stimulated the evolution ofethylene and acetaldehyde in male flowers predominantly, withresultant enhancement of the activity of phenylalanine ammonia-lyase(EC 4.3.1.5 [EC] ), and the browning of male flowers. A possible biochemicalpathway for browning of male flowers of Cryptomeria japonicais discussed. (Received September 19, 1994; Accepted September 7, 1995)     

Tyrosine phosphorylation of pp185 by insulin receptor kinase in a cell-free system

Insulin treatment of rat H-35 hepatoma cells causes rapid tyrosine phosphorylation of a high molecular weight protein termed pp185 besides autophosphorylation of the beta-subunit of the insulin receptor (IR) in an intact cell system. To elucidate the molecular basis for tyrosine phosphorylation of pp185, cell-free phosphorylation of pp185 was performed using phosphotyrosine-containing proteins (PYPs) purified from detergent-solubilized cell lysates by immunoprecipitation with anti-phosphotyrosine antibody. After insulin treatment of cells, marked increases of tyrosine phosphorylation of pp185 and IR were observed compared to noninsulin-treated cells. Site-specific antibodies that specifically inactivate IR kinase inhibited tyrosine phosphorylation of pp185 as well as the beta-subunit of IR. PYPs purified from detergent-free cell extracts contained pp185 but little IR; tyrosine phosphorylation of pp185 did not occur. Addition of IR kinase purified from human placenta to these PYPs restored insulin-dependent tyrosine phosphorylation of pp185. These results suggest that tyrosine phosphorylation of pp185 is catalyzed directly by IR kinase in this cell-free system.     

The discovery of YM-60828: a potent, selective and orally-bioavailable factor Xa inhibitor

Since Factor Xa (FXa) is well known to play a central role in thrombosis and hemostasis, inhibition of FXa is an attractive target for antithrombotic strategies. As a part of our investigation of a non-peptide, orally available FXa inhibitor, we found that a series of N-[(7-amidino-2-naphthyl)methyl]aniline derivatives possessed potent and selective inhibitory activities. Structure--activity relationship (SAR) of the substituent (R(1)) on the central aniline moiety suggested that increasing lipophilicity caused a detrimental effect on anticoagulant activity (prothrombin time assay) in plasma. Several compounds bearing a hydrophilic substituent in R(1) showed not only potent FXa inhibitory activities but also high anticoagulant activities. The best compound in this series was sulfamoylacetic acid derivative (YM-60828) which was a potent, selective and orally bioavailable FXa inhibitor and was chosen for clinical development.     

Localization of chemotactic activity and 64 kD protein phosphorylation for human polymorphonuclear leukocytes in N-terminus of the chemotactic protein LUCT/IL-8

A synthetic peptide, AVLPRSAKEL (LU10), the N-terminal amino acid sequence of chemotactic protein (LUCT/IL-8), showed chemotactic activity to polymorphonuclear leukocytes (PMN) with an ED50 of 5 nM for comparable to that of LUCT. Native LUCT and LU10 specifically induced the phosphorylation of 64 kD protein of PMN, and serine residue in the 64 kD protein was major phosphorylated amino acid. Furthermore, native LUCT enhanced the release of myeloperoxidase and beta-glucuronidase from PMN in the presence of cytochalasin B and FMLP, but LU10 did not. These results strongly suggest that the active site for both chemotactic stimulation and 64 kD protein phosphorylation is localized on the sequence of N-terminal 10 amino acids of LUCT.     

DEAD/H (Asp–Glu–Ala–Asp/His) box polypeptide 3, X-linked is an immunogenic target of cancer stem cells

Accumulating evidence suggests that most solid malignancies consist of heterogeneous tumor cells and that a relatively small subpopulation, which shares biological features with stem cells, survives through potentially lethal stresses such as chemotherapy and radiation treatment. Since the survival of this subpopulation of cancer stem cells (CSC) plays a critical role in recurrence, it must be eradicated in order to cure cancer. We previously reported that vaccination with CD133⁺ murine melanoma cells exhibiting biological CSC features induced CSC-specific effector T cells. These were capable of eradicating CD133⁺ tumor cells in vivo, thereby curing the parental tumor. In the current study, we indicated that DEAD/H (Asp–Glu–Ala–Asp/His) box polypeptide 3, X-linked (DDX3X) is an immunogenic protein preferentially expressed in CD133⁺ tumor cells. Vaccination with DDX3X primed specific T cells, resulting in protective and therapeutic antitumor immunity. The DDX3X-primed CD4⁺ T cells produced CD133⁺ tumor-specific IFNγ and IL-17 and mediated potent antitumor therapeutic efficacy. DDX3X is expressed in various human cancer cells, including lung, colon, and breast cancer cells. These results suggest that anti-DDX3X immunotherapy is a promising treatment option in efforts to eradicate CSC in the clinical setting.     

Supplemental effects and metabolic fate of crystalline arginine in juvenile shrimp Marsupenaeus japonicus

Feeding experiments with juvenile kuruma shrimp (Marsupenaeus japonicus) were conducted to understand the effects of supplemental levels of crystalline arginine hydrochloride on the growth and assimilation of arginine. In experiment 1 the juvenile shrimp were maintained on diets with and without arginine supplements. The addition of 3.0% arginine to a casein-based diet was slightly effective in improving the growth of the juveniles. In experiment 2, tracer experiments using [14C] arginine were conducted to clarify the ingestion and assimilation of arginine, 9 and 24 h after feeding, at different levels of supplemental arginine. Tracer experiments showed that the leaching rate of arginine 1 h after feeding ranged from 16 to 26% in the diets with different levels (0, 0.1, 1.5, 3.0 and 6.0%) of supplemental arginine; that is, 74-84% of the given arginine was actually ingested by the shrimp fed the diets. However, with increasing levels of supplemental arginine the incorporation rate of arginine into the whole body decreased and the utilization of absorbed arginine for body protein synthesis was reduced, whereas the excretion of absorbed arginine was increased. Thus, the absorbed arginine was not effectively utilized for body protein synthesis when large amounts of arginine were supplemented to the diets.