Lucigenin chemiluminescence assay as an adjunctive tool for assessment of various stages of inflammation: A study of quiescent inflammatory cells

A simple, fast, precise and biologically relevant toxicity assay for screening cytotoxicity of minerals would have distinct advantages due to its cost benefits and relative savings in time. Furthermore, a bioassay to differentiate acute and chronic in vivo pulmonary reactions could have potential value as predictors of fibrogenicity and pathogenicity. In this study we examined the potential use of lucigenin as a probe to evaluate the correlation between chemiluminescence (CL) generated by alveolar macrophages with the known cytotoxicity and patho genicity by conventional bioassays. In this study, we used small doses of dust (20 microg) to minimize cellular overload and to maintain homeostasis. Crystalline silica a highly fibrogenic dust was used as positive control and results are compared with those for bentonite, kaolin and talc. Among the three minerals compared with silica, bentonite was more reactive (27%) in CL assay and declined sharply compared to other minerals. This sudden decline in bentonite CL is caused by cytotoxicity leading to cell death. CL-induced by talc was comparable to silica and declines slowly. Kaolin on the other hand produced relatively a weaker (25%) CL compared to silica. Our data using relatively low doses of dust suggest that the CL assay may have a better predictive value in cytotoxicity evaluations compared to conventional toxicity assays.     

Applications of likelihood asymptotics for nonlinear regression in herbicide bioassays

Dose-response models are intensively used in herbicide bioassays. Despite recent advancements in the development of new herbicides, statistical analyses are commonly based on asymptotic approximations that are sometimes poor. This paper presents the use of recent results in higher order asymptotics for likelihood-based inference in nonlinear regression. The methods presented provide accurate approximation for the distribution of test statistics and for prediction limits. Analyses of the fit and measures of detection limits of the bioassays are considered, and the potential of the methods is illustrated by examples with real data.     

Pirimiphos-methyl resistance in two stored product mites, Acarus siro and Acarus farris, as detected by impregnated paper bioassay and esterase activity assays

The response to pirimiphos-methyl, in one strain of Acarus farris and two strains of Acarus siro, was assessed using an impregnated filter paper bioassay and by the selection of adults following exposure to pirimiphos-methyl. It was concluded that one of the strains of A. siro was resistant to pirimiphos-methyl and that a major resistance mechanism was involved. The second strain of A. siro gave a response similar to that of a laboratory strain unexposed to organophosphates and was considered to be susceptible. The A. farris strain responded to selection at the ED50 but not at the ED99, and it was concluded that a minor resistance mechanism is present in this strain. Assays of esterase activity were used to attempt to identify the biochemical mechanisms involved in the resistance detected by the bioassays. The A. farris and susceptible A. siro strains showed similar levels of esterase activity but the esterase activity of the resistant A. siro strain was significantly greater. An increase in esterase activity followed selection of both the A. farris strain and the resistant A. siro strain. An acetylcholinesterase assay showed no significant difference between the susceptible and pirimiphos-methyl selected strains of A. siro. The results suggest that esterases are involved in the resistance to pirimiphos-methyl found in A. siro and A. farris but that in A. siro, at least, other mechanisms may also be present.     

不同地区小菜蛾种群羧酸酯酶的毒理学性质研究

在1995～1997年对湖北武汉、河北张家口地区小菜蛾Plutella xylostella(L.)种群的抗药性进行了研究。结果表明对阿维菌素的抗性和台湾敏感种群相比,武汉种群抗性为4.3倍,张家口种群抗性为1.8倍;对马拉硫磷的抗性武汉和张家口种群分别为2.2和2.9倍;对氟铃脲的抗性分别为3.2和0.5倍;对溴氰菊酯的抗性分别为2.4和1.7倍。对羧酸酯酶(Care)的研究结果表明,三个种群幼虫CarE对a-乙酸萘酯或β-乙酸萘酯(a或β-NA)水解活性差异显著,但成虫Care活性没有明显差异。武汉和张家口种群幼虫CarE对a-NA和β-NA的亲和力没有明显差异,但是武汉种群幼虫Care对底物的亲和力高于张家口种群。敏感品系Care对a—NA的亲和力明显高于对β-NA,相差约3倍。不同类型的抑制剂对小菜蛾幼虫CarE的抑制能力不同。增效磷和对氧磷对敏感品系CarE水解a-NA具有明显的抑制作用,分别比对武汉种群Care的抑制作用大4.577倍(SVl)和2.576倍(对氧磷)。     

Toxicological analysis of the marine cyanobacterium Nodularia harveyana

Nodularia harveyana, a dinitrogen-fixing cyanobacterial isolate from the Mediteranean Sea, grown in an outdoor photobioreactor, was assayed for its bioactive compounds. The active substance(s) were lypophilic and showed strong allelopathic actvity against other axenic cyanobacteria, antibiotic activity against Gram-positive pathogenic bacteria and antifungal activity against two plant pathogens. The extracts were toxic (LC₅₀ at 30 μL) for grazers such as a rotifer (Brachionus calyciflorus) and a crustacean (Thamnocephalus platyurus). This revised version was published online in August 2006 with corrections to the Cover Date.     

Diamondback moth females oviposit more on plants infested by non‐parasitised than by parasitised conspecifics

Abstract 1. When offered a choice, female diamondback moths (Plutella xylostella) oviposited more eggs on plants with non‐parasitised conspecific larvae than on plants with parasitised larvae. 2. The leaf area consumed by parasitised larvae was significantly lower than that by non‐parasitised larvae. However, this quantitative difference in larval damage did not explain the female’s ability to discriminate between plants with parasitised and non‐parasitised larvae, as females showed an equal oviposition preference for plants infested by higher or lower densities of non‐parasitised larvae. 3. Pupal weight and duration of the larval stage of P. xylostella were independent of whether larvae were reared on plants that were previously infested by either non‐parasitised or parasitised larvae. 4. The larval parasitoid Cotesia vestalis did not distinguish between plants infested by non‐parasitised larvae and plants infested by larvae that had already been parasitised by conspecific wasps. 5. Based on these data, it can be concluded that the moth oviposition preference for plants infested by non‐parasitised conspecifics relative to plants infested by parasitised conspecifics was not explained by plant quality or by the attractiveness of plants towards wasps. It is hypothesised that one of the reasons for this preference is avoidance of plants where a relatively high risk of parasitism is expected due to the emergence of parasitoids from the parasitised host larvae.     

European populations of Diabrotica virgifera virgifera are resistant to aldrin,but not to methyl‐parathion

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a major pest of cultivated corn in North America and has recently begun to invade Europe. In addition to crop rotation, chemical control is an important option for D. v. virgifera management. However, resistance to chemical insecticides has evolved repeatedly in the USA. In Europe, chemical control strategies have yet to be harmonized and no surveys of insecticide resistance have been carried out. We investigated the resistance to methyl‐parathion and aldrin of samples from nine D. v. virgifera field populations originating from two European outbreaks thought to have originated from two independent introductions from North America. Diagnostic concentration bioassays revealed that all nine D. v. virgifera field populations were resistant to aldrin but susceptible to methyl‐parathion. Aldrin resistance was probably introduced independently, at least twice, from North America into Europe, as there is no evident selection pressure to account for an increase of frequency of aldrin resistance in each of the invasive outbreaks in Europe. Our results suggest that organophosphates, such as methyl‐parathion, may still provide effective control of both larval and adult D. v. virgifera in the European invasive outbreaks studied.     

Trade‐off between thermal tolerance and insecticide resistance in Plutella xylostella

Fitness costs associated with resistance to insecticides have been well documented, usually at normal temperature conditions, in many insect species. In this study, using chlorpyrifos‐resistant homozygote (R_R) and chlorpyrifos‐susceptible homozygote (S_S) of resistance ace1 allele of Plutella xylostella (DBM), we confirmed firstly that high temperature experience in pupal stage influenced phenotype of wing venation in insecticide‐resistant and insecticide‐susceptible Plutella xylostella, and S_S DBM showed significantly higher thermal tolerance and lower damages of wing veins under heat stress than R_R DBM. As compared to S_S DBM, R_R DBM displayed significantly lower AChE sensitivity to chlorpyrifos, higher basal GSTs activity and P450 production at 25°C, but higher inhibitions on the enzyme activities and P450 production as well as reduced resistance to chlorpyrifos under heat stress. Furthermore, R_R DBM displayed significantly higher basal expressions of hsp69s, hsp72s, hsp20, hsp90, Apaf‐1, and caspase‐7 at 25°C, but lower induced expressions of hsps and higher induced expressions of Apaf‐1, caspase‐9, and caspase‐7 under heat stress. These results suggest that fitness costs of chlorpyrifos resistance in DBM may partly attribute to excess consumption of energy caused by over production of detoxification enzymes and hsps when the proteins are less demanded at conducive environments but reduced expressions when they are highly demanded by the insects to combat environmental stresses, or to excess expressions of apoptotic genes under heat stress, which results in higher apoptosis. The evolutionary and ecological implications of these findings at global warming are discussed.     

家用调料植物粗提物防治小菜蛾的研究

以安全性和简易性为前提的庭园害虫防治成为休闲生活不可或缺的技术。本文以花椒Zanthoxylum bungeamtm、八角茴香lUicium veturn、辣椒Capsicum frutescens等日用品为材料,研究了不同种类、浓度和作用时间对小菜蛾Plutella xylostella这一重要庭园害虫的拒食、触杀及产卵忌避效果。拒食试验表明,供试3种植物对小菜蛾有显著拒食作用,拒食率从大到小依次为八角茴香（0．94）、花椒（0．86）、辣椒（0．76）;拒食率随处理时问长短变化的差异不显著,但随提取液浓度的升高而升高。触杀试验表明,供试3种植物对小菜蛾有显著触杀作用,24h内死亡率从大到小依次为花椒（45．6％）、八角茴香（35．6％）、辣椒（23．3％）,均显著高于对照处理的1．1％;触杀死亡率随虫龄的增加而降低。产卵忌避试验表明,3种植物对小菜蛾成虫产卵忌避作用由大到小依次为辣椒、八角茴香、花椒。综合本研究所得结果来看,八角茴香对小菜蛾的控制效果最好。所选材料容易获得,对小菜蛾忌避作用明显,在庭园小菜蛾防治中具有潜在的应用价值。     

Molecular cloning and expression profiles of calreticulin gene from the diamondback moth,Plutella xylostella

Calreticulin (CRT) plays pivotal roles in Ca²⁺ homeostasis, molecular chaperoning, infection, inflammation and innate immunity. In an attempt to study the involvement of CRT in innate immunity, the full-length cDNA of calreticulin (PxCRT) was cloned from the diamondback moth, Plutella xylostella. It consists of 1674 bp (excluding poly-A tail) with a longest open reading frame (ORF) of 1197 bp encoding 398 amino acids. In silico analysis of PxCRT ORF reveals that it has various repeat motifs and endoplasmic reticulum retention signal found in all the calreticulin proteins. As expected, high amino acid sequence identities were found from other CRTs identified from Bombyx mori (87%), Galleria mellonella (87%), Apis mellifera (74%), Anopheles gambiae (74%), Tribolium castaneum (73%), Culex quinquefasciatus (73%), Rhodnius prolixus (72%), Nasonia vitripennis (71%), Drosophila melanogaster (71%) and Haemaphysalis qinghaiensis (68%). During development, P. xylostella expressed PxCRT predominantly in the pupal stage. In addition, spatial expression pattern analysis indicates that PxCRT was highly expressed in the silk gland. PxCRT mRNA, furthermore, was strongly induced 3 to 6 h after laminarin treatment, suggesting that PxCRT appears to be involved in immune responses and also plays an important role in the silk gland.