Evaluation of partial sterility in mating performance and reproduction of the West Indian sweetpotato weevil,Euscepes postfasciatus

The sterile insect technique (SIT) is based on population and behavioral ecology and is widely used to suppress or eradicate target pest insect populations. The effectiveness of SIT depends on the ability of the released sterile males to mate with and inseminate wild females. The use of gamma‐radiation to induce sterility is, however, associated with negative impacts not only on reproductive cells but also on somatic cells. Consequently, irradiation for sterilization diminishes mating performance over time. In this study, we evaluated the balance between the irradiation dose and both fertility and mating propensity in Euscepes postfasciatus (Fairmaire) (Coleoptera: Curculionidae) for 22 days following irradiation. The mating propensity of males irradiated with a 150‐Gy dose, as currently used to induce complete sterility of E. postfasciatus in the SIT program in Okinawa Prefecture, was equal to that of non‐irradiated weevils for up to 6 days, and the mating propensity of males irradiated with a dose of 125 Gy was equal to that of non‐irradiated weevils for twice this period (12 days). The fertilization ability of weevils irradiated with a dose of 125 Gy was reduced by 4.6% in males and 0.6% in females, compared to the potential fertilization ability. We also discuss the possibility of the application of partially sterilized insects in eradication programs.     

Blebbistatin inhibits the chemotaxis of vascular smooth muscle cells by disrupting the myosin II-actin interaction

Blebbistatin is a myosin II-specific inhibitor. However, the mechanism and tissue specificity of the drug are not well understood. Blebbistatin blocked the chemotaxis of vascular smooth muscle cells (VSMCs) toward sphingosylphosphorylcholine (IC(50) = 26.1 +/- 0.2 and 27.5 +/- 0.5 microM for GbaSM-4 and A7r5 cells, respectively) and platelet-derived growth factor BB (IC(50) = 32.3 +/- 0.9 and 31.6 +/- 1.3 muM for GbaSM-4 and A7r5 cells, respectively) at similar concentrations. Immunofluorescence and fluorescent resonance energy transfer analysis indicated a blebbistatin-induced disruption of the actin-myosin interaction in VSMCs. Subsequent experiments indicated that blebbistatin inhibited the Mg(2+)-ATPase activity of the unphosphorylated (IC(50) = 12.6 +/- 1.6 and 4.3 +/- 0.5 microM for gizzard and bovine stomach, respectively) and phosphorylated (IC(50) = 15.0 +/- 0.6 microM for gizzard) forms of purified smooth muscle myosin II, suggesting a direct effect on myosin II motor activity. It was further observed that the Mg(2+)-ATPase activities of gizzard myosin II fragments, heavy meromyosin (IC(50) = 14.4 +/- 1.6 microM) and subfragment 1 (IC(50) = 5.5 +/- 0.4 microM), were also inhibited by blebbistatin. Assay by in vitro motility indicated that the inhibitory effect of blebbistatin was reversible. Electron-microscopic evaluation showed that blebbistatin induced a distinct conformational change (i.e., swelling) of the myosin II head. The results suggest that the site of blebbistatin action is within the S1 portion of smooth muscle myosin II.     

Effect of irradiation on mating performance and mating ability in the West Indian sweetpotato weevil, Euscepes postfasciatus

The sterile insect technique (SIT) is widely used for suppressing or eradicating target pest insect populations. The effectiveness of SIT depends on the ability of released sterile males to mate with and inseminate wild females. Irradiation not only damages the reproductive cells but the somatic cells as well. The mating behavior of irradiated males may be altered over time due to the depressed metabolic activity brought about by sterilization. In this study, we evaluated the mating behavior (copulation behavior, mating performance, and ability of sperm transfer) of irradiated males in Euscepes postfasciatus (Fairmaire) (Coleoptera: Curculionidae) for 16 days after irradiation in the laboratory. The mating performance of males irradiated with a 150 Gy dose, as currently used in the SIT program in Okinawa prefecture for E. postfasciatus, decreased compared to that of control after day 7. As a result, we considered that irradiation had no major effect on male mating behavior for approximately 1 week after irradiation.     

Structure of Malonogalactan: Carbon-13 Nuclear Magnetic Resonance Spectra of Malonogalactan

Six kinds of arachin subunits were isolated by isoelectric focusing in sucrose density gradient in the presence of 6 m urea and 0.2 m β-mercaptoethanol. These subunits (S₁ to S₆) had different isoelectric points (pI 5.8, 6.0, 6.3, 7.1, 7.4 and 8.3) and they were found in parent arachin in the weight ratio of 2.5 : 2.2 : 2.6 : 1.6 : 1.1 : 1.0. The molecular weights and N-terminal amino acids of the isolated subunits were 35,500 and valine (S₁), 37,500 and isoleucine (S₂), 40,500 and isoleucine (S₃) and 19,500 and glycine (S₄, S₅ and S₆). While, the parent arachin contained approximately one mole of valine, two moles of isoleucine and three moles of glycine as the N-terminal amino acids per 180,000 g. It is, therefore, concluded that arachin consists of six different subunits to form the molecular weight 180,000. No disulfide bond takes part in the subunit association of arachin, because the dissociation in a urea solution occurred in the absence of β-mercaptoethanol.     

Prolongation of the effective copulation period by fractionated‐dose irradiation in the sweet potato weevil,Cylas formicarius

The sterile insect technique (SIT), based on the principles of population and behavioral ecology, is widely used to suppress or eradicate target pest insect populations. The effectiveness of SIT depends on the ability of released sterile males to mate with and inseminate wild females; however, the use of gamma radiation to induce sterility negatively affects both somatic cells as well as reproductive cells. Consequently, sterilization by irradiation drastically diminishes mating performance over time. It is well known that fractionated‐dose irradiation, in which a sterilizing dose is delivered via a series of smaller irradiations, reduces radiation damage. In the present study, we evaluated the effect of fractionated‐dose irradiation on fertility, longevity, and mating propensity in Cylas formicarius (Summers) (Coleoptera: Brentidae) for 16 days after irradiation. Fractionated‐dose irradiation with 200 Gy induced full sterility regardless of the number of radiation doses. Although the mating propensity of males sterilized by a single 200 Gy dose (the current standard of the Okinawa Prefecture SIT program) was equal to that of non‐irradiated weevils for the first 6 days, the mating propensity of males sterilized by a series of three doses was maintained for at least the first 12 days. These results demonstrated that fractionated‐dose irradiation can be highly advantageous in C. formicarius eradication programs.     

Interaction between myosin and a trace amount of caldesmon

Caldesmon (CaD) is known as an actin binding protein. In this study, we proposed that a trace amount of caldesmon (TACD) could highly, efficiently, interact with myosin by producing a 'domino-like cascade' and characterized that TACD (lowest caldesmon/myosin molar ratio: 1/10,000) significantly increased precipitations and intrinsic tryptophan fluorescence intensity of myosin in both phosphorylated and unphosphorylated states compared to the base controls (P < 0.01). Actin-blocked TACD-myosin interaction, suggesting that it functioned as a negative regulator. Since CaD is not an enzyme, the in vivo significance of the highly efficient TACD-myosin interaction needs further investigation.     

Purification of a novel Ca-binding protein that inhibits myosin light chain kinase activity in lower eukaryote Physarum polycephalum

Myosin light chain kinase (MLCK) was partially purified from the lower eukaryote Physarum polycephalum. The activity to phosphorylate Physarum myosin was maximal in the absence of Ca2+ and decreased with an increase in Ca2+ concentration with a microM-level Kd. The Ca-binding protein contained in the MLCK preparation was purified to homogeneity. The native protein had a molecular mass of 75 kDa, while under denaturing conditions, it was 38 kDa. Ca-dependent changes in the intensities of intrinsic fluorescence showed that the Kd of the protein for Ca2+ was also in the microM-range. Our results suggest that the Ca-binding protein would play a key role in the effects of Ca2+ in the MLCK preparation.     

Irradiation does not affect field dispersal ability in the West Indian sweetpotato weevil, Euscepes postfasciatus

The effect of irradiation on the dispersal ability of males and females of the flightless West Indian sweetpotato weevil, Euscepes postfasciatus (Fairmaire) (Coleoptera: Curculionidae), a major sweet potato pest, was examined in the field using mark–release–recapture techniques. To evaluate the dispersal ability of the weevil, we released 7 619 weevils in two replicates (July and August 2007). Each replicate lasted 5 days from release to sampling and consisted of one weevil release and two weevil samplings. Thirty-two traps were placed in lines corresponding to eight compass directions and in four distance classes (8, 12, 16, and 20 m) in each replicate. We captured 709 (9.3%) weevils in the two replicates. Weevils dispersed at least 20 m from the release point in 2 days, regardless of sex or irradiation. Dispersal was strongly affected by wind direction, and in both replicates most weevils were recaptured in upwind directions. The mean dispersal distance for non-irradiated weevils was about 11 m per 2 days. Although there were some differences between sexes in recapture rate and dispersal distance, there was no consistent difference between irradiated and non-irradiated weevils in dispersal distance. We conclude that irradiation does not affect the dispersal ability of flightless E. postfasciatus in the field.