A simple method of monitoring for cypermethrin resistance in <Emphasis Type="Italic">Thrips tabaci</Emphasis> (Thysanoptera: Thripidae) using agar-coated glass pipettes

A simple plant-free method to monitor for cypermethrin resistance of Thrips tabaci Lindeman at 24 h after insect collection was developed, which utilizes an agar-coated glass pipette. In the laboratory, when cypermethrin-resistant and -susceptible insects were mixed in various ratios and subjected to the method, only resistant insects were detected as survivors. All survivors in the field bioassay were found to have a particular amino acid mutation (T929I) in the sodium channel. Results obtained in this study also showed that the method is affected by temperature, possibly because pyrethroid toxicity increases as the temperature decreases. This method could be used for the on-site monitoring of T. tabaci for cypermethrin resistance with careful temperature management after insect collection.     

Effect of irradiation on dispersal ability of male sweetpotato weevils (Coleoptera: Brentidae) in the field

We used the mark-and-recapture method in the field to test the effect of gamma radiation on the dispersal ability of the male sweetpotato weevil, Cylas formicarius elegantulus (Summers) (Coleoptera: Brentidae), a serious sweetpotato pest in Japan. To evaluate the dispersal ability of male sweetpotato weevil, we released 27,218 males (13,302 males irradiated with a dose of 200 Gy and 13,916 nonirradiated males) in two replications (September and October 2005). Each replication lasted 5 d from the release of weevils to the removal of traps, and male weevils were released twice (1 and 3 d before trap setting). Forty pheromone traps were placed in lines corresponding to eight compass directions and five distance classes (20, 50,100, 200, and 500 m) in each replication. We captured 2,263 irradiated males (17.0%) and 2,007 nonirradiated males (14.4%) in the two replications. Six irradiated and eight nonirradiated males were captured in the traps 500 m far from the release point. All parameters to evaluate the dispersal ability of irradiated male sweetpotato weevil (recapture rate, dispersal distance, and dispersal direction) were similar to nonirradiated males in three of the four trials. However, parameters were different between irradiated males and nonirradiated males in one trial. Because the majority of parameters consistently show that the similarity of the dispersal ability, we considered that male sweetpotato weevil irradiated with a dose of 200 Gy possessed equal dispersal ability to that of nonirradiated males in the field.     

A 1-Hour Enzyme-Linked Immunosorbent Assay for Quantitation of Acrolein- and Hydroxynonenal-Modified Proteins by Epitope-Bound Casein Matrix Method

A simple and rapid enzyme-linked immunosorbent assay (ELISA) method for quantitation of acrolein and 4-hydroxy-2-nonenal (HNE)-modified proteins was developed. Microtiter plate wells were precoated and blocked simultaneously with epitope-bound bovine caseins as matrix proteins, and aldehyde-modified proteins were quantitated by a competition assay with a monoclonal antibody specific for acrolein-modified lysine or HNE-modified histidine epitopes. Minimal reaction times required for the coating/blocking; first monoclonal antibody and the peroxidase-conjugated second antibody binding steps were 3, 3, and 7 min, respectively, the former two steps being found to be or akin to diffusion-rate-limiting reactions. The convenient ELISA should find an application for analyses of the intricate processes involved in oxidative stress and carcinogenic insult. The epitope-attachment methodology may also be advantageous for the quantitation of various other biologically important haptenic molecules.     

In vitro system for high-throughput screening of random peptide libraries for antimicrobial peptides that recognize bacterial membranes.

Antibacterial peptides have been isolated from a wide range of species. Some of these peptides act on microbial membranes, disrupting their barrier function. With the increasing development of antibiotic resistance by bacteria, these antibacterial peptides, which have a new mode of action, have attracted interest as antibacterial agents. To date, however, few effective high-throughput approaches have been developed for designing and screening peptides that act selectively on microbial membranes. In vitro display techniques are powerful tools to select biologically functional peptides from peptide libraries. Here, we used the ribosome display system to form peptide-ribosome-mRNA complexes in vitro from nucleotides encoding a peptide library, as well as immobilized model membranes, to select specific sequences that recognize bacterial membranes. This combination of ribosome display and immobilized model membranes was effective as an in vitro high-throughput screening system and enabled us to identify motif sequences (ALR, KVL) that selectively recognized the bacterial membrane. Owing to host toxicity, it was not possible to enrich any sequence expected to show antimicrobial activity using another in vitro system, e.g. phage display. The synthetic peptides designed from these enriched motifs acted selectively on the bacterial model membrane and showed antibacterial activity. Moreover, the motif sequence conferred selectivity onto native peptides lacking selectivity, and decreased mammalian cell toxicity of native peptides without decreasing their antibacterial activity.     

Studies on freshwater red algae of Malaysia III. development of carposporophytes ofBatrachospermum cayennense Montagne,B. beraense Kumano andB. hypogynum Kumano et Ratnasabapathy

The development of the carposporophytes ofBatrachospermum cayennense, B. beraense andB. hypogynum is observed. The carpogonium bearing branch ofB. cayennense is very long and its cells are differentiated from those of the vegetative fascicles in size and shape. The trichogyne nucleus ofB. cayennense andB. beraense is not recognized. In the above two species it is observed that after contact with the trichogyne the nucleus of the spermatium divides into two daughter nuclei, one of which fuses with the female nucleus, while the other remains in the trichogyne. InB. beraense the fertilized carpogonium is divided transversely and the gonimoblast filaments are produced from the divided carpogonium. The protoplasmic connection between the carpogonium and the hypogynous nutritive cell is especially pronounced inB. hypogynum. Carpospores ofB. hypogynum germinate within the carposporangia and young germlings often throw away original carpospores, hence these young germlings look like gemmae.     

Detection and Characterization of Antibodies to Bacterial Heat-Shock Protein 60 in Sera of Patients with Primary Biliary Cirrhosis

The enzyme-linked immunosorbent assay (ELISA) with bacterial heat-shock protein 60 (HSP60) purified from Yersinia enterocolitica (Ye) revealed that the antibodies directed against YeHSP60 existed in sera of patients with primary biliary cirrhosis (PBC). To characterize the epitope specificity of the antibodies in patients, the epitope mapping of HSP60 by means of the antibodies was performed. The results have suggested that the epitope recognized with anti-HSP60 antibodies in PBC relates to the amino acid sequence of YeHSP60 molecule as follows: DLGQA-KRVVINKDTTIIIDGVGDEAAIQGRLAQIRQQIEEATSDYDKEK.     

The autoregulation of nodulation mechanism is related to leaf development

To understand the autoregulation of nodulation (AON) system, in which leguminous plants control the nodule number, we examined the details of the characteristics of hypernodulation soybean mutants NOD1-3 and NOD3-7. A microscopic study showed that NOD1-3 and NOD3-7 produced small-size leaves due to the smaller number of leaf cells, compared with the Williams parent. These phenotypes were not affected by inoculation with bradyrhizobia or nitrate supply. The AON signaling might be related to the control system of leaf cell proliferation. This hypothesis was strongly supported by the finding that activation of AON in wild types by inoculation leads to an increase in the cell number of leaves.     

Effect of irradiation on mating ability in the male sweetpotato weevil (Coleoptera: Curculionidae)

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 is the effective manner to sterilize mass-reared insects. The negative impacts of this procedure are not limited to damage on reproductive cells. Gamma-radiation damages the epithelial tissue of midgut, which affects the alimentation in insects. Irradiated males alter their mating behavior over time because of the depression of metabolic activity by sterilization. In this study, we evaluated the male mating performance and sexually compatibility of irradiated male Cylas formicarius elegantulus (Summers) (Coleoptera: Curculionidae) with a 200-Gy dose, as currently used in the SIT program in Okinawa Prefecture, throughout 16 d after irradiation in the laboratory. The mating ability of irradiated males did not differ from that of control males for about a week. However, the mating ability of irradiated male drastically decreased thereafter. We consider that irradiated male C. formicarius elegantulus with a 200-Gy dose had no major effect on male mating behavior approximately for a week after irradiation.