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Plants release volatiles in response to caterpillar feeding that attract natural enemies of the herbivores, a tri-trophic interaction which has been considered an indirect plant defence against herbivores. The caterpillar-induced plant volatiles have been reported to repel or attract conspecific adult herbivores. To date however, no volatile signals that either repel or attract conspecific adults under field conditions have been chemically identified. Apple seedlings uniquely released seven compounds including acetic acid, acetic anhydride, benzyl alcohol, benzyl nitrile, indole, 2-phenylethanol, and (E)-nerolidol only when infested by larvae of the light brown apple moth, Epiphyas postvittana. In field tests in New Zealand, a blend of two of these, benzyl nitrile and acetic acid, attracted a large number of conspecific male and female adult moths. In North America, male and female adults of the tortricid, oblique-banded leafroller, Choristoneura rosaceana, were most attracted to a blend of 2-phenylethanol and acetic acid. Both sexes of the eye-spotted bud moth, Spilonota ocellana, were highly attracted to a blend of benzyl nitrile and acetic acid. This study provides the first identification of caterpillar-induced plant volatiles that attract conspecific adult herbivores under natural conditions, challenging the expectation of herbivore avoidance of these induced volatiles.

The polycomb repressive complex 2 (PRC2) methylates lysine 27 of histone H3 (H3K27) through its catalytic subunit Ezh2. PRC2-mediated di- and tri-methylation (H3K27me2/H3K27me3) have been interchangeably associated with gene repression. However, it remains unclear whether these two degrees of H3K27 methylation have different functions. In this study, we have generated isogenic mouse embryonic stem cells (ESCs) with a modified H3K27me2/H3K27me3 ratio. Our findings document dynamic developmental control in the genomic distribution of H3K27me2 and H3K27me3 at regulatory regions in ESCs. They also reveal that modifying the ratio of H3K27me2 and H3K27me3 is sufficient for the acquisition and repression of defined cell lineage transcriptional programs and phenotypes and influences induction of the ESC ground state., (Published by Elsevier Inc.)

Background: Drosophila suzukii is a major pest of cherry in the western United States. We evaluated whether the addition of sugary baits could improve the efficacy of two classes of insecticides not considered to be sufficiently effective for this pest, diamides and spinosyns, in laboratory and field trials in cherry., Results: Adding cane sugar alone or in combination with the yeasts Saccharomyces cerevisiae or Aureobasidium pullulans significantly improved insecticide efficacy. However, the significance of adding yeasts to the sugar plus insecticide on fly mortality varied with respect to both the insecticide and yeast species. The addition of S. cerevisiae to sugar also did not significantly reduce egg densities in fruit compared with sugar alone. The addition of a yeast plus sugar significantly reduced egg densities in three field trials with cyantraniliprole and in two out of three trials with spinosad., Conclusion: The addition of cane sugar with or without yeast can improve the effectiveness of diamide and spinosyn insecticides for D. suzukii in cherry. Inclusion of these two insecticides in D. suzukii management programs may alleviate the strong selection pressure currently being imposed on a few mode-of-action insecticide classes used by growers to maintain fly suppression over long continuous harvest periods of mixed cultivars. Published 2015. This article is a U.S. Government work and is in the public domain in the USA., (Published 2015. This article is a U.S. Government work and is in the public domain in the USA.)

Field studies were conducted in the United States, Hungary, and New Zealand to evaluate the effectiveness of septa lures loaded with ethyl (E,Z)-2,4-decadienoate (pear ester) and (E)-4,8-dimethyl-1,3,7-nonatriene (nonatriene) alone and in combination with an acetic acid co-lure for both sexes of codling moth, Cydia pomonella (L.). Additional studies were conducted to evaluate these host plant volatiles and acetic acid in combination with the sex pheromone, (E,E)-8,10-dodecadien-1-ol (codlemone). Traps baited with pear ester/nonatriene + acetic acid placed within orchards treated either with codlemone dispensers or left untreated caught significantly more males, females, and total moths than similar traps baited with pear ester + acetic acid in some assays. Similarly, traps baited with codlemone/pear ester/nonatriene + acetic acid caught significantly greater numbers of moths than traps with codlemone/pear ester + acetic acid lures in some assays in orchards treated with combinational dispensers (dispensers loaded with codlemone/pear ester). These data suggest that monitoring of codling moth can be marginally improved in orchards under variable management plans using a binary host plant volatile lure in combination with codlemone and acetic acid. These results are likely to be most significant in orchards treated with combinational dispensers. Significant increases in the catch of female codling moths in traps with the binary host plant volatile blend plus acetic acid should be useful in developing more effective mass trapping strategies., (Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.)

Studies were conducted with the codling moth granulosis virus (CpGV) to evaluate whether adding the yeast Saccharomyces cerevisiae Meyen ex E. C. Hansen with brown cane sugar could improve larval control of Cydia pomonella (L.). Larval mortalities in dipped-apple bioassays with S. cerevisiae or sugar alone were not significantly different from the water control. The addition of S. cerevisiae but not sugar with CpGV significantly increased larval mortality compared with CpGV alone. The combination of S. cerevisiae and sugar with CpGV significantly increased larval mortality compared with CpGV plus either additive alone. The addition of S. cerevisiae improved the efficacy of CpGV similarly to the use of the yeast Metschnikowia pulcherrima (isolated from field-collected larvae). The proportion of uninjured fruit in field trials was significantly increased with the addition of S. cerevisiae and sugar to CpGV compared with CpGV alone only in year 1, and from the controls in both years. In comparison, larval mortality was significantly increased in both years with the addition of S. cerevisiae and sugar with CpGV compared with CpGV alone or from the controls. The numbers of overwintering larvae on trees was significantly reduced from the control following a seasonal program of CpGV plus S. cerevisiae and sugar. The addition of a microencapsulated formulation of pear ester did not improve the performance of CpGV or CpGV plus S. cerevisiae and sugar. These data suggest that yeasts can enhance the effectiveness of the biological control agent CpGV, in managing and maintaining codling moth at low densities., (Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.)

The response of Grapholita molesta (Busck) males to three-component sex pheromone blends containing a 100% ratio of the major sex pheromone component, (Z)-8-dodecenyl acetate and a 10% ratio of (Z)-8-dodecenol, but with varying ratios of (E)-8-dodecenyl acetate (0.4, 5.4, 10.4, 30.4, and 100.1% E-blends) was tested with populations in eight stone and pome fruit orchards in Europe, Asia, and North and South America. Traps baited with the 5.4% E-blend caught significantly more males than traps with any other blend with all populations. Significantly more males were caught in traps baited with the 10.4% E-blend than in traps with the remaining blends, except with the 0.4% E-blend in Turkey. Significant differences in male moth catches occurred between the other blends with the 0.4>30.4% E-blend, and the 30.4>100.1% E-blend. Male moth catches with the 100.1% E-blend only differed from the hexane control in Chile. No apparent differences were noted to these blends in populations collected from pome or stone fruits. Flight tunnel assays to synthetic blends with a subset of populations were similar to the field results, but the breadth of the most attractive E-blends was wider. Flight tunnel assays also demonstrated a high level of male-female cross-attraction among field-collected populations. Female gland extracts from field-collected populations did not show any significant variation in their three-component blends. The only exceptions in these assays were that long-term laboratory populations were less responsive and attractive, and produced different blend ratios of the two minor components than recently collected field populations.

Studies were conducted to assess glacial acetic acid (GAA) with various host plant volatiles (HPVs) and the sex pheromone, (E,E)-8, 10-dodecadien-1-ol, of codling moth, Cydia pomonella (L), as lures in traps for tortricid pests that often co-occur in tree fruits in the western United States. In addition to codling moth, field trapping studies were conducted with oriental fruit moth, Grapholita molesta (Busck), obliquebanded leafroller Choristoneura rosaceana (Harris), the leafroller Pandemis pyrusana Kearfott, and the eyespotted budmoth, Spilonota ocellana (Denis and Schiffermüller). HPVs included ethyl (E,Z)-2,4-decadienoate (pear ester), (E)-4,8-dimethyl-1,3,7-nonatriene, butyl hexanoate, (E)-β-ocimene, (E)-β-farnesene, and farnesol. Three types of GAA co-lures differing in a 10-fold range in weekly evaporation rates were tested. The evaporation rate of GAA co-lures was an important factor affecting moth catches. The highest rate tested captured fewer codling moth but more leafrollers and eyespotted budmoth. GAA co-lures caught both sexes of each species. The field life of butyl hexanoate and (E)-β-ocimene lures were much shorter than pear ester or sex pheromone lures. Adding GAA to pear ester or to (E)-β-ocimene significantly increased the catches of only codling moth or oriental fruit moth, respectively. Combining pear ester or (E)-β-ocimene with GAA did not affect the catch of either species compared with the single more attractive HPV. Adding HPVs to GAA did not increase the catches of either leafroller species or eyespotted budmoth. Traps baited with pear ester, sex pheromone, and GAA for monitoring codling moth were also effective in classifying pest pressure of both leafroller species within orchards.

Neurodegenerative diseases represent an increasing burden in our aging society, yet the underlying metabolic factors influencing onset and progression remain poorly defined. The relationship between impaired IGF-1/insulin-like signaling (IIS) and lifespan extension represents an opportunity to investigate the interface of metabolism with age-associated neurodegeneration. Using data sets of established DAF-2/IIS-signaling components in Caenorhabditis elegans, we conducted systematic RNAi screens in worms to select for daf-2-associated genetic modifiers of α-synuclein misfolding and dopaminergic neurodegeneration, two clinical hallmarks of Parkinson's disease. An outcome of this strategy was the identification of GPI-1/GPI, an enzyme in glucose metabolism, as a daf-2-regulated modifier that acts independent of the downstream cytoprotective transcription factor DAF-16/FOXO to modulate neuroprotection. Subsequent mechanistic analyses using Drosophila and mouse primary neuron cultures further validated the conserved nature of GPI neuroprotection from α-synuclein proteotoxicity. Collectively, these results support glucose metabolism as a conserved functional node at the intersection of proteostasis and neurodegeneration., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The genetic structure of adult codling moth, Cydia pomonella (L.), populations was characterized both inside a managed apple, Malus domestica Borkdhausen, orchard and in surrounding unmanaged hosts and nonhost trees in central Chile during 2006-2007. Adult males were collected using an array of sex pheromone-baited traps. Five microsatellite genetic markers were used to study the population genetic structure across both spatial (1-100 ha) and temporal (generations within a season) gradients. Analysis of molecular variance (AMOVA) found a significant, but weak, association in both the spatial and temporal genetic structures. Discriminant analysis also found significant differentiation between the first and second generation for traps located either inside or outside the managed orchard. The Bayesian assignment test detected three genetic clusters during each of the two generations, which corresponded to different areas within the unmanaged and managed apple orchard interface. The lack of a strong spatial structure at a local scale was hypothesized to be because of active adult movement between the managed and unmanaged hosts and the asymmetry in the insecticide selection pressure inside and outside the managed habitats. These data highlight the importance of developing area-wide management programs that incorporate management tactics effective at the landscape level for successful codling moth control.

Parkinson's disease (PD) is a neurodegenerative brain disorder characterized by selective dopaminergic (DAergic) cell loss that results in overt motor and cognitive deficits. Current treatment options exist to combat PD symptomatology, but are unable to directly target its pathogenesis due to a lack of knowledge concerning its etiology. Several genes have been linked to PD, including three genes associated with an early-onset familial form: parkin, pink1 and dj1. All three genes are implicated in regulating oxidative stress pathways. Another hallmark of PD pathophysiology is Lewy body deposition, associated with the gain-of-function genetic risk factor α-synuclein. The function of α-synuclein is poorly understood, as it shows both neurotoxic and neuroprotective activities in PD. Using the genetically tractable invertebrate Caenorhabditis elegans (C. elegans) model system, the neurotoxic or neuroprotective role of α-synuclein upon acute Mn exposure in the background of mutated pdr1, pink1 or djr1.1 was examined. The pdr1 and djr1.1 mutants showed enhanced Mn accumulation and oxidative stress that was reduced by α-synuclein. Moreover, DAergic neurodegeneration, while unchanged with Mn exposure, returned to wild-type (WT) levels for pdr1, but not djr1.1 mutants expressing α-synuclein. Taken together, this study uncovers a novel, neuroprotective role for WT human α-synuclein in attenuating Mn-induced toxicity in the background of PD-associated genes, and further supports the role of extracellular dopamine in exacerbating Mn neurotoxicity.

Mitochondria are cellular power plants that supply ATP to power various biological activities essential for neuronal growth, survival, and function. Due to extremely varied morphological features, neurons face exceptional challenges to maintain energy homeostasis. Neurons require specialized mechanisms distributing mitochondria to distal synapses where energy is in high demand. Axons and synapses undergo activity-dependent remodeling, thereby altering mitochondrial distribution. The uniform microtubule polarity has made axons particularly useful for exploring mechanisms regulating mitochondrial transport. Mitochondria alter their motility under stress conditions or when their integrity is impaired. Therefore, research into the mechanisms regulating mitochondrial motility in healthy and diseased neurons is an important emerging frontier in neurobiology. In this chapter, we discuss the current protocols in the characterization of axonal mitochondrial transport in primary neuron cultures isolated from embryonic rats and adult mice. We also briefly discuss new procedures developed in our lab in analyzing mitochondrial motility patterns at presynaptic terminals and evaluate their impact on synaptic vesicle release.

The combination of a pathogenic virus and mutualistic yeasts isolated from larvae of codling moth Cydia pomonella is proposed as a novel insect control technique. Apples were treated with codling moth granulovirus (CpGV) and either one of three yeasts, Metschnikowia pulcherrima, Cryptococcus tephrensis, or Aureobasidium pullulans. The combination of yeasts with CpGV significantly increased mortality of neonate codling moth larvae, compared with CpGV alone. The three yeasts were equally efficient in enhancing the activity of CpGV. The addition of brown cane sugar to yeast further increased larval mortality and the protection of fruit against larvae. In comparison, without yeast, the addition of sugar to CpGV did not produce a significant effect. A field trial confirmed that fruit injury and larval survival were significantly reduced when apple trees were sprayed with CpGV, M. pulcherrima, and sugar. We have shown earlier that mutualistic yeasts are an essential part of codling moth larval diet. The finding that yeast also enhances larval ingestion of an insect-pathogenic virus is an opportunity for the development of a novel plant protection technique. We expect the combination of yeasts and insect pathogens to essentially contribute to future insect management.

Codling moth, Cydia pomonella L. (Lepidoptera: Tortricidae), is a major cosmopolitan pest of apple and other pome fruits. Ethyl (E,Z)-2,4-decadienoate (pear ester) has been identified as a host-derived kairomone for female and male codling moths. However, pear ester has not performed similarly in different fruit production areas in terms of the relative magnitude of moth catch, especially the proportion of females caught. Our work was undertaken to identify host volatiles from apples, and to investigate whether these volatiles can be used to enhance the efficacy of host kairomone pear ester for monitoring female and male codling moths. Volatiles from immature apple trees were collected in the field using dynamic headspace sampling during the active period of codling moth flight. Using gas chromatography-electroantennogram detector (GC/EAD) analysis, six compounds elicited responses from antennae of females. These compounds were identified by GC/mass spectrometry (MS) and comparisons to authentic standards as nonanal, (E)-4,8-dimethyl-1,3,7-nonatriene, methyl salicylate, decanal, (Z,E)-α-farnesene, and (E,E)-α-farnesene. When the EAD-active compounds were tested individually in the field, no codling moths were caught except for a single male with decanal. However, addition of (E)-4,8-dimethyl-1,3,7-nonatriene, methyl salicylate, decanal, or (E,E)-α-farnesene to pear ester in a binary mixture enhanced the efficacy of pear ester for attracting female codling moths compared to pear ester alone. Addition of the 6-component blend to the pear ester resulted in a significant increase in the number of males attracted, and enhanced the females captured compared to pear ester alone; the number of males and females caught was similar to that with the pear ester plus acetic acid combination lure. Our results demonstrate that it is possible to synergize the response of codling moth to host kairomone by using other host volatiles. The new apple-pear ester host kairomone blend should be helpful for monitoring female codling moth, and may provide the basis for further improvement of codling moth kairomone.

Oil-coated clear panel traps baited with a host plant-based kairomone lure have successfully been used for monitoring female grape berry moth, Paralobesia viteana (Clemens) (Lepidoptera: Tortricidae), but low capture rates as well as difficulty in servicing these traps makes them unsuitable for commercial use. We compared the performance of different trap designs in a flight tunnel and in a vineyard by using a 7-component synthetic kairomone blend, with a focus on trap visual cues. In flight tunnel experiments, a clear delta trap performed better than other traps. When we tested clear delta, green delta, or clear wing traps baited with a cut grape shoot, >50% of female grape berry moths made complete upwind flights. However, the clear delta trap was the only design that resulted in female moths entering the trap. Similar results were observed when females were tested with different traps (clear delta, green delta, white delta, clear wing, or green wing traps) baited with the kairomone lure. Adding a visual pattern that mimicked grape shoots to the outside surface of the clear delta trap resulted in 66% of the females that made upwind flights entering the trap. However, the positive effect of adding a visual pattern to the trap was not observed in a vineyard setting, where clear delta traps with or without a visual pattern caught similar numbers of females. Still, the number of male and female grape berry moths captured in clear delta traps with or without a visual pattern was not significantly different from the number of male and female grape berry moths captured in panel traps, suggesting that the use of these delta traps could be a less cumbersome alternative to oil-coated panel traps for monitoring female grape berry moth.

Background: The possibility of improving the efficacy of various insecticides for codling moth, Cydia pomonella (L.), by the addition of a microencapsulated formulation of pear ester, ethyl (2E, 4Z)-2,4-decadienoate (PE-MEC, 5% AI), was evaluated in field trials in apple from 2005 to 2009., Results: The addition of PE-MEC (<3.0 g AI ha(-1) ) significantly lowered fruit injury with low rates of organophosphate and neonicotinoid insecticides. The addition of PE-MEC (1.48 g AI ha(-1) ) to maximum label rates of insecticides with moderate activity for codling moth, such as ebamectin benzoate, spinosad and methoxyfenozide, significantly improved their effectiveness. However, PE-MEC did not consistently improve the activity of maximum label rates of either rynaxypyr or spinetoram. The addition of PE-MEC with seasonal spray programs of acetamiprid and azinphos-methyl in combination with the use of sex pheromones for mating disruption provided the most effective control., Conclusion: The addition of PE-MEC can improve the effectiveness of some insecticides for codling moth in apple. Insecticides active via dermal toxicity are more strongly improved by the addition of PE-MEC. Season-long use of PE-MEC can also augment the effectiveness of mating disruption. Adding PE-MEC to variable integrated management programs for C. pomonella should be further investigated., (This article is a US Government work and is in the public domain in the USA. Published 2012 by John Wiley & Sons, Ltd.)

Purpose: With a theoretical link between stent insertion and increased risk of tumour seeding, there is concern about long-term survival after the use of self-expanding metallic stents (SEMS) as a "bridge to surgery" in the treatment of left-sided obstructing colorectal cancer. This cohort study aims to determine if preoperative stenting adversely affects long-term survival by comparing a group of patients having preoperative stenting (group A) with a group of patients having elective surgery (group B) in a single centre., Methods: The study is retrospective. Survival was calculated with Kaplan-Meier analysis and compared using the log-rank test. Other group characteristics were compared with Fisher's exact test., Results: From November 1998 to November 2008, 15 patients had preoperative SEMS and were entered in group A. This represented 11.5 % of a total of 130 patients undergoing SEMS insertion in the same period. Group B included 88 consecutive patients undergoing elective left-sided colonic resection for Dukes' B and C cancer excluding mid and low rectal tumours between January 2003 and December 2007. The 30-day mortality rate for groups A and B was 6.7 % (one patient) and 5.7 % (five patients), respectively. The 5-year survival rate was 60 % and 58 %, respectively, with a p value of 0.96., Conclusions: In our own practice, patients undergoing SEMS as a "bridge to surgery" have the same long-term survival with those undergoing elective surgery. This finding needs to be confirmed in larger scale studies.

Lysosomes are responsible for degradation and recycling of bulky cell material, including accumulated misfolded proteins and dysfunctional organelles. Increasing evidence implicates lysosomal dysfunction in several neurodegenerative disorders, including Parkinson's disease and related synucleinopathies, which are characterized by the accumulation of α-synuclein (α-syn) in Lewy bodies. Studies of lysosomal proteins linked to neurodegenerative disorders present an opportunity to uncover specific molecular mechanisms and pathways that contribute to neurodegeneration. Loss-of-function mutations in a lysosomal protein, ATP13A2 (PARK9), cause Kufor-Rakeb syndrome that is characterized by early-onset parkinsonism, pyramidal degeneration and dementia. While loss of ATP13A2 function plays a role in α-syn misfolding and toxicity, the normal function of ATP13A2 in the brain remains largely unknown. Here, we performed a screen to identify ATP13A2 interacting partners, as a first step toward elucidating its function. Utilizing a split-ubiquitin membrane yeast two-hybrid system that was developed to identify interacting partners of full-length integral membrane proteins, we identified 43 novel interactors that primarily implicate ATP13A2 in cellular processes such as endoplasmic reticulum (ER) translocation, ER-to-Golgi trafficking and vesicular transport and fusion. We showed that a subset of these interactors modified α-syn aggregation and α-syn-mediated degeneration of dopaminergic neurons in Caenorhabditis elegans, further suggesting that ATP13A2 and α-syn are functionally linked in neurodegeneration. These results implicate ATP13A2 in vesicular trafficking and provide a platform for further studies of ATP13A2 in neurodegeneration.

Traps baited with ethyl (E,Z)-2,4-decadienoate (pear ester) or (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) in two- or three-way combinations with the sex pheromone (E,E)-8,10-dodecadien-1-ol (codlemone) and acetic acid (AA) were evaluated for codling moth, Cydia pomonella (L.). All studies were conducted in apple orchards, Malus domestica Borkhausen, treated with sex pheromone dispensers during 2010. Septa were loaded with codlemone, DMNT, and pear ester individually or codlemone with either DMNT or pear ester together (combo lures). Polyethylene vials loaded with AA were added as a co-lure. Residual analyses of field-aged combo lures and weight loss of the AA co-lure were conducted. AA vials lost 50-150 mg wk(-1). Weekly weight loss was not affected by field aging, but was closely correlated with the daily mean temperature. Pear ester was released from septa at a slightly higher but nonsignificant rate than codlemone. DMNT was released at a significantly higher rate than codlemone, and lures were effective for 4 wk. The addition of codlemone to traps baited with either host plant volatile plus AA had either no effect or significantly increased total moth catches. The addition of AA significantly increased the catch of female moths with either combo lure. Total moth catches in traps baited with pear ester or DMNT combo lures and AA did not differ and were either significantly higher or similar to the pear ester combo lure. These data suggest that codling moth may be more effectively monitored in sex pheromone-treated orchards with multi-component lures, including codlemone, AA, and host plant volatiles.

The efficacy of combining insecticides with a microencapsulated formulation of ethyl (2E,4Z) -2,4-decadienoate (pear ester, PE-MEC) was evaluated in walnuts, Juglans regia L., for codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), and navel orangeworm, Amyelois transitella Walker (Lepidoptera, Pyralidae). Two types of studies were conducted to compare the use of insecticides with and without PE-MEC. In the first study, PE-MEC in combination with reduced rates of insecticides, including chlorpyrifos, phosmet, methoxyfenozide, and codling moth granulovirus were evaluated in single tree replicates. PE-MEC was tested at one to three rates (0.6, 1.8, and 4.4 g active ingredient ha(-1)) with each insecticide. In the second study, seasonal programs including sprays of esfenvalerate, chlorpyrifos, and ethyl parathion at full rates were evaluated in replicated two ha blocks. Significant reductions in nut injury occurred in the single-tree trial with treatments of PE-MEC plus insecticide compared with the insecticides used alone against both pest species; except with methoxyfenozide for navel orangeworm. Similarly, nut injury in the large plots was significantly reduced with the addition of PE-MEC, except for navel orangeworm in one of the two studies. These results suggest that adding pear ester as a microencapsulated spray can improve the efficacy of a range of insecticides for two key pests and foster the development of integrated pest management tactics with reduced insecticide use in walnut.

Parkinson's disease (PD), an adult neurodegenerative disorder, has been clinically linked to the lysosomal storage disorder Gaucher disease (GD), but the mechanistic connection is not known. Here, we show that functional loss of GD-linked glucocerebrosidase (GCase) in primary cultures or human iPS neurons compromises lysosomal protein degradation, causes accumulation of α-synuclein (α-syn), and results in neurotoxicity through aggregation-dependent mechanisms. Glucosylceramide (GlcCer), the GCase substrate, directly influenced amyloid formation of purified α-syn by stabilizing soluble oligomeric intermediates. We further demonstrate that α-syn inhibits the lysosomal activity of normal GCase in neurons and idiopathic PD brain, suggesting that GCase depletion contributes to the pathogenesis of sporadic synucleinopathies. These findings suggest that the bidirectional effect of α-syn and GCase forms a positive feedback loop that may lead to a self-propagating disease. Therefore, improved targeting of GCase to lysosomes may represent a specific therapeutic approach for PD and other synucleinopathies., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Protein misfolding and aggregation are key pathological features observed in numerous neurodegenerative diseases, including the misfolding of α-synuclein (α-syn) in Parkinson's disease (PD) and β-amyloid in Alzheimer's disease. While this phenomenon is widely observed, the etiology and progression of these diseases is not fully understood. Furthermore, there is a lack of therapeutic treatments directed at halting the progression and neurodegeneration associated with these diseases. This demands a need for an inexpensive, easy to manipulate multicellular organism to conduct both genetic and chemical screens within to identify factors that may play a pivotal role in the pathology of these diseases. Herein, we describe methodology involved in identifying genetic modifiers of α-syn misfolding and toxicity in the nematode roundworm, Caenorhabditis elegans. Transgenic nematodes engineered to express human α-syn in the body wall muscles or dopaminergic (DA) neurons result in formation of cytoplasmic puncta or DA neurodegeneration, respectively. Using these models, we describe the use of RNA interference (RNAi) and transgenic gene expression to functionally elucidate potential therapeutic gene targets that alter α-syn misfolding and DA neurotoxicity., (Copyright © 2010 Elsevier Inc. All rights reserved.)

We analyzed the spatial distribution and dispersal of codling moth, Cydia pomonella (L.), adults within two heterogeneous agroecosystems typical of central Chile: commercial apple, Malus domestica Borkhausen, orchards surrounded by various unmanaged host plants. Both a geostatistical analysis of catches of adult males with a grid of sex pheromone-baited traps and an immunological self-marking technique combined with traps baited with a male and female attractant were used. The spatial analyses identified the key sources of moths within these diverse landscapes. Codling moth catches in traps were spatially associated within distances of ≈ 150-300 m. Similarly, the mean distance from the immunological self-marking plots within the commercial apple orchard to the traps that captured marked adults was 282 m. In contrast, the mean distance in the capture of marked moths from unmanaged self-marking plots to a commercial orchard was 828 m. These data suggest that the success of any future area-wide management programs for codling moth in Chilean pome fruit must include a component for managing or removing noncommercial hosts that surround orchards. This analysis also suggests that the selection pressure for resistance imposed by insecticide sprays within managed orchards is likely dampened by the influx of susceptible moths from unmanaged sites common in central Chile.

Background: Codling moth, Cydia pomonella (L.), has been intensely managed with the organophosphate insecticide azinphos-methyl for 50 years, and populations have developed resistance. New management programs have been developed and implemented that rely more heavily on other classes of insecticides. A prerequisite for developing effective resistance management strategies for these compounds is to establish their current levels of effectiveness. Adult and neonate larval assays were conducted to assess the response of field-collected codling moth populations from apple in Washington State., Results: Male codling moth populations exhibited a range of responses to a discriminating concentration of azinphos-methyl in a survey of 20 populations. Populations from certified organic orchards were more susceptible than those from conventional orchards. Mean fecundity was inversely related to azinphos-methyl tolerance. Male responses to azinphos-methyl and acetamiprid varied significantly among populations and were correlated. The residual effectiveness of field applications of both insecticides varied significantly against neonate larvae. Neonate bioassays with insecticide-dipped fruit found significant differences among populations with azinphos-methyl, acetamiprid, methoxyfenozide and spinosad, but not with esfenvalerate., Conclusion: These results support a concern that alternation of insecticides with different modes of action may not be a sufficient strategy to avoid the evolution of broad-spectrum insecticide resistance by codling moth., (Published 2010 by John Wiley & Sons, Ltd.)

Background: Studies surveyed the toxicity of several insecticides against adult codling moth, Cydia pomonella (L.), and examined the field effectiveness of applying low-volume (12 L ha(-1)) sprays alone or in combination with a microencapsulated (MEC) sex pheromone formulation., Results: Neonicotinyls, organophosphates and synthetic pyrethroids significantly reduced fecundity at concentrations nearly 100-fold lower than their maximum labeled field rate. Field studies in 2005 demonstrated that six applications of esfenvalerate resulted in > 90% reduction in fruit injury versus the untreated check. The addition of the MEC pheromone formulation did not further improve control. Five sprays of esfenvalerate, phosmet and acetamiprid all significantly reduced levels of fruit injury compared with the untreated control in 2006. Esfenvalerate and acetamiprid mixed with the MEC pheromone significantly reduced fruit injury compared with the MEC-only treatment. Significant increases in pest and decreases in predator mite densities occurred in plots treated with esfenvalerate in both years. Low-volume sprays of phosmet and acetamiprid did not disrupt mites., Conclusion: Low-volume insecticide sprays can effectively manage codling moth and are less disruptive of integrated mite management. Developing an effective 'attract and kill' technology with this approach will require optimization of the attractant(s) to maximize moth exposure to insecticide residues., (Published 2010 by John Wiley & Sons, Ltd.)

The relative performance of a clear delta trap baited with individual or combination sex pheromone and kairomone lures for codling moth, Cydia pomonella L., was evaluated against three vertical clear pane and colored delta traps in field trials within apple, Malus domestica (Borkhausen). The clear caught significantly more moths of each sex than an orange delta trap when baited with ethyl (E,Z)-2,4-decadienoate (pear ester) and acetic acid (PE+AA) and performed similarly to oil-coated pane traps. The clear caught significantly more females than the orange delta trap with pear ester alone, pear ester plus (E,E)-8,10-dodecadien-1-ol (codlemone) (PE-PH), or a combination of PE-PH and acetic acid. Male moth capture was similar in both clear and colored delta traps with all codlemone lures tested. Seasonal moth catches of female moths were higher in clear than either white (second flight, 2008) or orange (first and second flights, 2009) traps baited with PE-PH. Total moth catch was significantly higher in clear than white traps in 2008 and did not differ between clear and orange traps in 2009. Clear traps baited with acetic acid but not with pear ester, PE-PH, or when unbaited caught significantly more nontarget moths than colored traps. These studies suggest that the use of clear traps with their higher captures of female codling moths could improve both monitoring programs and the development of lure and kill strategies.

Studies were conducted to examine the deposition of microcapsules and the attractiveness of treated apple leaves, Malus domestica Borkhausen, for codling moth, Cydia pomonella L., after low-volume concentrated sprays [24.7 g (AI) in 12 liters of water/ha] of a microencapsulated (MEC) sex pheromone formulation (CheckMate CM-F). Nearly 30% of leaves collected from sprayed zones within tree canopies had no microcapsules, whereas 20% had >20 microcapsules. Microcapsule density was correlated with leaf area, and significant differences in the density of microcapsules per leaf were found because of both height and depth in the canopy relative to the sprayed zone and leaf surface. In general, the highest concentration of microcapsules was deposited on the underside of leaves in the sprayed zone. However, deposition was greater on the upper than the bottom surface of leaves in the canopy below the spray zone and in the tops of trees on the opposite side of the canopy. Field-aged MEC-treated apple leaves elicited upwind flight and moth contact in flight tunnel tests for at least 5 wk. Precipitation reduced the attractiveness of leaves, particularly for leaves treated only on their upper versus bottom surface. Traps in unsprayed orchards baited with MEC-treated artificial leaves were attractive for 5 wk. Moth catches in similar traps placed in MEC-sprayed plots were low but increased significantly over 3-4 wk. These data suggest that, after a brief initial period of sensory disruption, low-volume MEC sprays create point sources of sex pheromone (leaves) within the orchard that may enhance mating disruption through competitive attraction.

Transgenic Caenorhabditis elegans can be readily created via microinjection of a DNA plasmid solution into the gonad. The plasmid DNA rearranges to form extrachromosomal concatamers that are stably inherited, though not with the same efficiency as actual chromosome. A gene of interest is co-injected with an obvious phenotypic marker, such as rol-6 or GFP, to allow selection of transgenic animals under a dissecting microscope. The exogenous gene may be expressed from its native promoter for cellular localization studies. Alternatively, the transgene can be driven by a different tissue-specific promoter to assess the role of the gene product in that particular cell or tissue. This technique efficiently drives gene expression in all tissues of C. elegans except for the germline or early embryo. Creation of transgenic animals is widely utilized for a range of experimental paradigms. This video demonstrates the microinjection procedure to generate transgenic worms. Furthermore, selection and maintenance of stable transgenic C. elegans lines is described.

The application of a microencapsulated (MEC) sex pheromone formulation (Checkmate CM-F) for codling moth, Cydia pomonella (L.), in low volume, concentrated sprays was evaluated in a series of small plot and grower trials in apple, Malus domestica Borkhausen, and pear, Pyrus communis L. Preliminary tests found that MEC sprays applied at 172-207 kilopascals in 12-23 liters/ha deposited the highest density of microcapsules per leaf. The addition of a latex sicker did not increase the deposition of microcapsules. Small plot tests in 2004 compared the effectiveness of two low-volume sprayers against a standard high-volume spray (926 liters/ha) applied at 1,379 kilopascals. Moth catches and fruit injury were significantly lower in plots treated with the low-volume sprays compared with plots treated with the standard sprayer. These results suggest that concentrating the MEC formulation increases the deposition of microcapsules and improves its effectiveness. Larger trials were conducted with a low-volume sprayer in 4-ha plots within commercial apple (2005-2006) and pear orchards (2005) paired with similar plots treated with hand-applied pheromone dispensers. Levels of fruit injury were not significantly different between pheromone treatments in any of the three tests. Moth catches, however, were significantly higher in the MEC- versus the dispenser-treated apple plots in 2005. No difference was found in the fruit injury levels in MEC-treated apple orchards in 2005 caused by irrigation method, but moth catches were significantly higher in overhead versus undertree orchards. The advantages and current limitations of using MEC sex pheromone sprays to supplement current grower's management strategies for codling moth is discussed.

Improvements to the diagnosis and treatment of Parkinson's disease (PD) are dependent upon knowledge about susceptibility factors that render populations at risk. In the process of attempting to identify novel genetic factors associated with PD, scientists have generated many lists of candidate genes, polymorphisms, and proteins that represent important advances, but these leads remain mechanistically undefined. Our work is aimed toward significantly narrowing such lists by exploiting the advantages of a simple animal model system. While humans have billions of neurons, the microscopic roundworm Caenorhabditis elegans has precisely 302, of which only eight produce dopamine (DA) in hemaphrodites. Expression of a human gene encoding the PD-associated protein, alpha-synuclein, in C. elegans DA neurons results in dosage and age-dependent neurodegeneration. Worms expressing human alpha-synuclein in DA neurons are isogenic and express both GFP and human alpha-synuclein under the DA transporter promoter (Pdat-1). The presence of GFP serves as a readily visualized marker for following DA neurodegeneration in these animals. We initially demonstrated that alpha-synuclein-induced DA neurodegeneration could be rescued in these animals by torsinA, a protein with molecular chaperone activity. Further, candidate PD-related genes identified in our lab via large-scale RNAi screening efforts using an alpha-synuclein misfolding assay were then over-expressed in C. elegans DA neurons. We determined that five of seven genes tested represented significant candidate modulators of PD as they rescued alpha-synuclein-induced DA neurodegeneration. Additionally, the Lindquist Lab (this issue of JoVE) has performed yeast screens whereby alpha-synuclein-dependent toxicity is used as a readout for genes that can enhance or suppress cytotoxicity. We subsequently examined the yeast candidate genes in our C. elegans alpha-synuclein-induced neurodegeneration assay and successfully validated many of these targets. Our methodology involves generation of a C. elegans DA neuron-specific expression vector using recombinational cloning of candidate gene cDNAs under control of the Pdat-1 promoter. These plasmids are then microinjected in wild-type (N2) worms, along with a selectable marker for successful transformation. Multiple stable transgenic lines producing the candidate protein in DA neurons are obtained and then independently crossed into the alpha-synuclein degenerative strain and assessed for neurodegeneration, at both the animal and individual neuron level, over the course of aging.

Genomic multiplication of the locus-encoding human alpha-synuclein (alpha-syn), a polypeptide with a propensity toward intracellular misfolding, results in Parkinson's disease (PD). Here we report the results from systematic screening of nearly 900 candidate genetic targets, prioritized by bioinformatic associations to existing PD genes and pathways, via RNAi knockdown. Depletion of 20 gene products reproducibly enhanced misfolding of alpha-syn over the course of aging in the nematode Caenorhabditis elegans. Subsequent functional analysis of seven positive targets revealed five previously unreported gene products that significantly protect against age- and dose-dependent alpha-syn-induced degeneration in the dopamine neurons of transgenic worms. These include two trafficking proteins, a conserved cellular scaffold-type protein that modulates G protein signaling, a protein of unknown function, and one gene reported to cause neurodegeneration in knockout mice. These data represent putative genetic susceptibility loci and potential therapeutic targets for PD, a movement disorder affecting approximately 2% of the population over 65 years of age.

Studies were conducted with codling moth, Cydia pomonella L., to fit cumulative curves for the occurrence of injured fruits and male moth catches in sex pheromone-baited traps as a function of accumulated degree-days after the start of moth flight. Twelve data sets were collected from seven apple, Malus domestica Bordhausen, orchards in Washington State from 2003 to 2006. Cumulative data were grouped across years for orchards either treated with sex pheromone dispensers or untreated and fit to logistic equations for both the first and second generation. No significant differences were found for the cumulative curves of moth flight or egg hatch between pheromone-treated and untreated orchards; thus, these data were combined. These new logistic models for moth flight and egg hatch were compared with a widely used distributed-delay model originally developed in Michigan. The cumulative flight curves for the logistic and distributed-delay models were statistically different (slopes) for the first but not the second generation. Cumulative egg hatch in the logistic model was significantly different from the distributed-delay model (intercepts and slopes) for both generations. Most strikingly, the timing of 50% egg hatch during the first generation was delayed 100 DD in the logistic model. The potential impact of this change in the characterization of codling moth's phenology on the effectiveness of insecticide programs targeting eggs and newly eclosed larvae was examined. Possible explanations for this significant difference between the models are discussed.

(E,Z) -2,4-decadienoate (pear ester) is a larval kairomone for the codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae). Orchard studies were conducted in 2005 and 2006 in apple, Malus domestica Borkhausen, and pear, Pyrus communis L., to evaluate a 5% active ingredient (AI), microencapsulated formulation of pear ester (PE-MEC) as an insecticidal additive for the codling moth granulovirus (CpGV). Although CpGV applied at 5-15-d intervals at commercial rates (2.2 X 10(12)-10(13) granules per ha) killed the majority (82-94%) of larvae found inside infested fruit, it did not eliminate significant damage, i.e., 30-92% fruit injury at harvest versus 51-82% in controls. PE-MEC treatments had significant but inconsistent results in our tests. In apple (mixed cultivars), PE-MEC (3.7-4.7 g [AI] /ha) plus CpGV reduced the percentage of fruit injured during the second but not the first larval generation, compared with CpGV alone, but there no was no additional population reduction (live larvae collected from infested fruit and tree bands). In 'Bartlett' pear, PE-MEC (3.7 g [AI] /ha) plus CpGV significantly increased larval mortality and reduced deep fruit entries at harvest over CpGV alone in 2006, but similar improvements were not observed in 2005 when a lower rate (1.5 g [AI] /ha) was tested. Surprisingly, compared with untreated controls, the PE-MEC formulation alone also reduced fruit injury (mid-season in Bartlett) and larval survivorship inside infested fruit at harvest (2006 apple tests and both years in Bartlett). Although pear ester seems amenable as a kairomonal adjuvant for use with insecticides, our inconsistent data with CpGV in apple and pear suggest practical improvements in formulation and application strategies (e.g., to optimize and maintain attractive release rates) are needed.

The influence of trap placement on catches of codling moth, Cydia pomonella L., was examined in a series of studies conducted in orchards treated with Isomate-C Plus sex pheromone dispensers. Mark-recapture tests with sterilized moths released along the interface of pairs of treated and untreated apple and pear plots found that significantly more male but not female moths were recaptured on interception traps placed in the treated plots. In a second test, significantly higher numbers of wild male and female moths were caught on interception traps placed in treated versus untreated plots within a heavily infested orchard. The highest numbers of male moths were caught on traps placed along the interior edge of the treated plots. Trap position had no influence on the captures of female moths. In a third test, north-south transects of sex pheromone-baited traps were placed through adjacent treated and untreated plots that received a uniform release of sterilized moths. Traps on the upwind edge of the treated plots caught similar numbers of moths as traps upwind from the treated plots. Moth catch was significantly reduced at all other locations inside versus outside of the treated plots, including traps placed on the downwind edge of the treated plot. In a fourth test, five apple orchards were monitored with groups of sex pheromone-baited traps placed either on the border or at three distances inside the orchards. The highest moth counts were in traps placed at the border, and the lowest moth counts were in traps placed 30 and 50 m from the border. In a fifth test, the proportion of traps failing to catch any moths despite the occurrence of local fruit injury was significantly higher in traps placed 50 versus 25 m from the border. The implications provided by these data for designing an effective monitoring program for codling moth in sex pheromone-treated orchards are discussed.

Studies were conducted with codling moth, Cydia pomonella L., to evaluate the mating status of male and female moths in apple, Malus domestica (Borkhausen), orchards treated with and without sex pheromone dispensers. Laboratory studies first examined the effect of multiple mating of male and female moths on female fecundity and egg fertility. Females that had mated three times had a significantly higher fecundity than singly mated moths. Sequential mating by male moths had no effect on the fecundity of female moths or egg fertility. However, male moth age did impact female fecundity, with significantly fewer eggs laid after mating with virgin 1- versus 3-d-old males. The mean size of the first spermatophore transferred by males was significantly larger than all subsequent spermatophores. Classifying spermatophores based on size was used in field sampling to categorize the mating status of the female's partner. The proportion of mated females with small spermatophores (partner had previously mated) was significantly higher in treated versus untreated orchards. The proportion of female moths caught in traps baited with pear ester that were virgin was low (

The influence of the anthranilic diamide insecticide chlorantranilipole (DPX-E2Y45; Rynaxypyr against the adult stage of codling moth, Cydia pomonella L., was examined. Insecticide residues in plastic cups sprayed with 56 mg AI L(-1) (equivalent to the recommended field rate) had a minimal effect on adult survival or fecundity, but significantly fewer female moths were mated in treated than in untreated cups. Similar results were found in field studies with adults placed in screened cages on treated apple foliage for up to 3 weeks. Mating was disrupted when either sex was topically dosed with technical material (0.01-0.1 microg). Males were more sensitive to the disruptive effects of chlorantranilipole than females. A 2 h exposure significantly reduced male activity (walking and wing fanning), but not female calling. A 25 h exposure significantly reduced the activity of both sexes. The proportion of males landing near a sex pheromone source in a flight tunnel was significantly reduced following exposures of 2-4 h and completely eliminated following a 17 h exposure. Male moths partially recovered from either topical or residual exposure to chlorantranilipole, but the extent of their recovery was dose and time dependent. Male response to sex pheromone recovered within 2 h following a 4 h exposure, but following a 17 h exposure only a partial recovery occurred after 48 h. Mating by moth pairs exposed to chlorantranilipole residues for 24 h did not significantly increase during recovery periods of 24-96 h, except in cups treated with 10% of the standard concentration. The proportion of field-collected female codling moths caught in pear ester-baited traps that were mated was significantly lower in apple orchards treated season-long with chlorantranilipole compared with females caught in either untreated orchards or those treated with sex pheromones to cause mating disruption.

The rainfastness of a microencapsulated sex pheromone formulation for codling moth, Cydia pomonella (L.), was evaluated in a series of laboratory experiments with detached apple, pear, and walnut leaves. Increasing the intensity and duration of simulated rainfall significantly increased the removal of microcapsules from both the top and bottom of apple leaves. The removal of microcapsules was significantly higher from the top versus the bottom of leaves at all rates tested. Leaf angle was a significant factor affecting the removal of microcapsules from the top surface of apple leaves with fewer microcapsules removed, because leaves were oriented with a steeper downward angle. Both leaf surfaces of apple and pear retained a higher proportion of microcapsules than walnut leaves, and the bottom surface of apple leaves retained significantly more than pear leaves. Three spray adjuvants were evaluated as stickers for microcapsules. No difference was found in the number of microcapsules deposited on apple leaves among three stickers tested at rates from 0.06 to 0.25%. However, in a second test a latex sticker significantly increased the deposition of microcapsules on apple leaves compared with a polyvinyl polymer and a pine resin sticker at a rate of 0.06%. Significantly more microcapsules were retained on the bottom versus the top of apple leaves with all stickers. The latex and polyvinyl stickers significantly increased the retention of microcapsules versus the pine resin sticker and the control on apple leaves. In another test, the addition of 0.06% latex sticker did not increase the deposition of microcapsules on any of the three leaf types. However, the addition of the latex sticker significantly increased the retention of microcapsules on the top of apple and pear leaves and the bottom of apple leaves. The addition of a latex sticker did not affect the retention of microcapsules on walnut leaves.

This multi-author paper reviews current work by USDA-ARS scientists in the field of chemical ecology. Work with pheromones, the discovery and development of the codling moth kairomone, studies on insect-plant interactions and chemically mediated tritrophic plant-insect interactions have led to practical methods for control of important insect pests.

The alkyl ethyl and methyl esters of (2E,4Z)-2,4-decadienoic acid found in head-space samples of ripe Bartlett pear (Pyrus communis L.) stimulated a response from neonate larvae of the codling moth (CM), Cydia pomonella (L.), in both static-air Petri-plate and in upwind Y-tube and straight-tube olfactometer bioassays. In comparison with the known CM neonate attractant, (E,E)-alpha-farnesene, ethyl (2E,4Z)-2,4-decadienoate was attractive at 10-fold and 1,000-fold lower threshold dosages in the Petri-plate and in the Y-tube bioassays, respectively. Methyl (2E,4Z)-2,4-decadienoate was attractive to CM neonates in these bioassays at much higher doses than ethyl (2E,4Z)-2,4-decadienoate. Other principal head-space volatiles from ripe pear fruit and pear leaves, including butyl acetate, hexyl acetate, (Z)-3-hexenyl acetate, and (E)-beta-ocimene, were not attractive to CM neonates. The potential uses of these pear kairomones for monitoring and control of CM in walnuts and apple are discussed.

Ethyl (2E, 4Z)-2,4-decadienoate, a pear-derived volatile, is a species-specific, durable, and highly potent attractant to the codling moth (CM), Cydia pomonella (L.), a serious pest of walnuts, apples, and pears worldwide. This kairomone attracts both CM males and virgin and mated females. It is highly attractive to CM in both walnut and apple orchard contexts, but has shown limited effectiveness in a pear orchard context. Rubber septa lures loaded with ethyl (2E, 4Z)-2,4-decadienoate remained attractive for several months under field conditions. At the same low microgram load rates on septa, the combined gender capture of CM in kairomone-baited traps was similar to the capture rate of males in traps baited with codlemone, the major sex pheromone component. The particular attribute of attracting CM females renders this kairomone a novel tool for monitoring population flight and mating-ovipositional status, and potentially a major new weapon for directly controlling CM populations.

A diet-incorporation larval bioassay was developed to measure the response of codling moth, Cydia pomonella (L.), to the benzoylhydrazine insecticides tebufenozide and methoxyfenozide. The bioassay tested neonates and third, fourth, and fifth instars from a laboratory colony and neonates and fourth instars from a pooled population collected from five certified-organic apple orchards. Bioassays were scored after 6 and 14 d. No differences between the laboratory and field population were found for either insecticide. Significant differences were found in the response of third and fifth instars between the 6 and 14 d bioassays, primarily due to a high proportion of moribund larvae in the shorter assay. Larval age had a significant effect in bioassays and was more pronounced in 6- versus 14-d tests. Fifth instars were significantly less susceptible to both insecticides than other stages, while responses of third and fourth instars were similar. The response of neonates was significantly different from third and fourth instars to tebufenozide but not with methoxyfenozide in the 14-d test. Field bioassays excluded the use of fifth instars and were scored after 14 d. LC50s estimated for 18 field-collected populations varied five- and ninefold for tebufenozide and methoxyfenozide, respectively. The responses of all but six field-collected populations were significantly different from the laboratory strain. Five of these six populations were collected from orchards with no history of organophosphate insecticide use. The LC50 for methoxyfenozide of one field-collected population reared in the laboratory for three generations declined fourfold, but was still significantly different from the laboratory population. These data suggest that transforming current codling moth management programs in Washington from a reliance on organophosphate insecticides to benzoylhydrazines may be difficult.

Male and female codling moth, Cydia pomonella (L.), were monitored with passive interception traps (PI-traps) in apple orchards treated with sex pheromone dispensers. The proportion of mated females recaptured by PI-traps was significantly higher than the proportion released after the release of both sexes into a codling moth-infested orchard. However, no significant difference occurred between the proportion of mated females recaptured and released when only females were released into uninfested orchards. Replicated nine-tree apple plots situated either on the edge or in the center ofpheromone-treated apple orchards were monitored with PI-traps during first moth flight in 1995 and during both flights in 1996. Moths caught on PI-traps were predominately males. The first male moths were captured 7-10 d before females during the first flight in both years. Initial capture of virgin and mated females on PI-traps coincided in 1995. Mated females were captured 14 d after the first virgin females in 1996. The mean proportion of females that were mated ranged from 32 to 55% during the first flight and 85 to 92% during the second flight. Moth catch and fruit injury were significantly higher in the edge versus the center plots. The numbers of total and female moths caught with PI-traps were significantly correlated with fruit injury for each generation. The percentage of female moths caught on PI-traps that were mated was 32% lower and the mean oocyte load of all females was 42% higher in a pheromone-treated apple orchard than in the untreated crabapple grove monitored during May and June 1997.

The effectiveness of tebufenozide applied against the adult, egg, and larvae of codling moth, Cydia pomonella (L.), was evaluated. Significant reductions in fecundity and egg hatch occurred after 1-h and 24-h exposures of females and 24-h exposures of males-only to residues in plastic plates. A significant reduction in egg hatch was also found after a 1-h exposure of males. The ovicidal effects of tebufenozide in field trials did not significantly differ for eggs laid on residues or treated topically. Corrected egg mortality exceeded 95% for cohorts laid <130 degree-days after sprays were applied. Fecundity and egg hatch were measured after either a 24-h exposure of moths or a 10-d exposure of moths and eggs on apple trees. Significant reductions in both fecundity and fertility occurred compared with an untreated control up to 7 d in the 24-h assays and accounted for 60-70% fewer larvae produced per mated female. The mean numbers of larvae produced per mated female after the 10-d exposure were reduced 100-75% in assays started on day 0-21 after the spray application compared with the untreated control. Fecundity was significantly reduced for 7 d and egg hatch was reduced for the entire 21-d test period in these trials. Residues had a 14-d half-life on apple foliage. Residues applied to foliage or to foliage and fruit did not significantly increase the proportion of uninjured fruit compared with the untreated control in bioassays where neonates were placed on foliage 10 cm from fruit. However, the proportion of injured fruits with shallow stings versus deep entries was higher on the treated versus the untreated plants. Field applications of tebufenozide reduced fruit injury >90% when applied early during emergence. Plots treated after the beginning of egg hatch had a larger proportion of injured fruits with shallow larval feeding.

Studies were conducted in 1997 to evaluate the effects of the kaolin-based particle film formulation M96-018 on adults, eggs, and larvae of the obliquebanded leafroller, Choristoneura rosaceana (Harris). Particle film treatments significantly reduced female longevity, mating success, and number of egg masses oviposited compared with moths on untreated apple leaves in sleeve-cage and screen-cage tests. No differences in mating success or oviposition were caused by the application rates and coverage density of M96-018 on foliage. Females avoided ovipositing on particle film-treated leaves in choice tests. Larval hatch was not affected by topical application or residual exposure to M96-018. Larval weight gain and pupal weight were significantly reduced and larval mortality increased in no-choice feeding tests with M96-018. In choice tests, larvae preferred to feed on untreated leaf surfaces. The negative effects on larval development and survivorship on M96-018-treated foliage did not differ across a fourfold difference in spray application rate. A significant reduction in the number of infested shoots was found in orchard trials when M96-018 was applied before bud break in late March compared with untreated trees. No reductions in larval densities were found compared with an untreated control following prebloom and postbloom applications.

Studies were conducted in 1997 and 1998 to evaluate the effects of three particle film formulations consisting of kaolin and adjuvants on neonate larvae, ovipositing adult females, and eggs of the codling moth, Cydia pomonella (L.). Neonate larval walking speed, fruit discovery rate, and fruit penetration rate on apple host plants coated with particle films were significantly lower than on host plants without particle films in laboratory assays. Females oviposited less on host plants covered with a particle film residue than on untreated plants in laboratory choice and no-choice tests. Hatch rate of codling moth neonate larvae was unaffected by particle films sprayed on host plants either before or after oviposition. Fruit infestation rates were significantly reduced on particle film-treated trees compared with untreated trees for both first- and second-generation codling moth in field trials in both apple and pear orchards. Particle films appear to be a promising supplemental control approach for codling moth in orchards where moth density is high, and may represent a stand-alone method where moth densities are lower.

Introduction: Reduced vascular compliance in patients with hypertension results from an increase in extra-cellular matrix (ECM) protein deposition in blood vessels. At least two key factors, namely mechanical strain and neurohumoral mediators, for example Angiotensin II (Ang II), promote fibrogenesis within vessel walls; however potential interactions between these have not been clearly defined. This work examined the direct effect of mechanical strain on matrix mRNA expression and protein synthesis by human vascular smooth muscle (VSM) cells and identified the importance of renin-angiotensin system (RAS) activation in stretch-induced matrix production., Methods: Human VSM cells were exposed either to a cyclical mechanical strain regimen or to Ang II in the presence or absence of the Ang II receptor (AT(1) R) antagonist losartan or its more potent metabolite EXP3174. Analysis of matrix mRNA expression (Northerns) and protein synthesis (ELISA) and cellular AT(1)-receptor protein expression (Westerns) were determined., Results: Ang II increased both collagen alpha1 (92%, SEM +/- 20%) mRNA expression and fibronectin (21% +/- 6%) protein synthesis in static VSM cells compared with unstimulated controls. The effect of Ang II was attenuated by antagonism of the AT(1)-receptor (AT(1) R). Similarly, mechanical strain induced an increase in both collagen alpha1 (102% +/- 30%) mRNA expression and fibronectin (50% +/-21%) protein synthesis. Surprisingly, in the absence of exogenous Ang II, AT(1)-receptor blockade attenuated this stretch-induced increase in matrix synthesis. Mechanical strain also induced an increase in total cellular AT(1)-receptor protein (30.7% +/- 3.5%) compared with static cells., Conclusion: Both mechanical strain and Ang II increased matrix gene expression and protein synthesis by human VSM cells. The effect of strain was attenuated by AT(1)-receptor antagonism. Our results further suggest that mechanical strain may sensitise human VSM cells to the fibrogenic actions of Ang II, perhaps via upregulation of the AT(1)-receptor.

Background: There have been few studies documenting the optimal cervical cytologic technique in pregnancy. This study was initiated to investigate the effectiveness and safety of two standard techniques., Methods: Two hundred twenty-two new obstetrical patients at a family practice residency program signed informed consent, were enrolled, and were randomized for cervical cytologic screening using either Zelsmyr Cytobrush and Ayre spatula or Dacron swab and Ayre spatula. The pathologists were blinded to the study, and results were reported with standard Bethesda System nomenclature. Patients were asked about complications at follow-up visits. Statistical analysis was performed using Fisher's exact test and the two-sample t test, with statistical significance set at P < .05., Results: The Cytobrush-spatula technique yielded 95.6% (108/113) smears with adequate endocervical cells, as compared with 69.7% (76/109) of smears obtained with the Dacron swab and spatula (P = < .0001, odds ratio 9.38). Cytologic atypia, defined as noninflammatory cellular abnormalities, was uncovered on 15.9% (18/113) of the smears obtained with the Cytobrush and spatula technique and on 13.8% (15/109) of the smears obtained with the Dacron swab and spatula (P = .7082). The complication rate, which included spot bleeding and spontaneous abortion, occurred in 6.5% (7/108) of the smears obtained with the Cytobrush and 3.8% (4/105) of the smears obtained with the Dacron swab (P = .54)., Conclusions: The Cytobrush-spatula technique significantly increased endocervical cell yield, did not increase detection of cytologic atypia, and did not increase the risk of adverse maternal or fetal outcomes when used for cervical cytologic screening during pregnancy. These results support the use of the Cytobrush-spatula technique over the swab and spatula for obtaining cervical cytologic smears in pregnancy.