Structure and behaviour of the sperm terminal filament

Light- and electron-microscopic observations of Ciona and Lytechinus spermatozoa show a thin terminal filament at the distal end. The terminal filament is 5-6 microns long and contains the two central microtubules and a variable number of A-tubule extensions of the peripheral doublet microtubules. The transition from the 9 + 2 region to the terminal filament is tapered more gradually in Lytechinus than in Ciona. Photographs of the movement of beating spermatozoa do not show any obvious discontinuity in curvature at the transition region. Bends are propagated smoothly off the end of the flagellum with no decrease in curvature. However, spermatozoa in which the terminal filament has been removed show a clear 'end effect'. This end effect involves a rapid unbending of bends that have reached the distal end of the flagellum. Computer simulations of flagellar models lacking a terminal filament show a similar end effect. Addition of a terminal filament to the end of the computer model can eliminate the end effect. Realistic bending behaviour of the model is obtained by using a terminal filament with a tapered elastic bending resistance in the basal portion of the terminal filament and a value of 0.03 x 10^(9) pN nm^2 in the remainder of the terminal filament. This leads to estimates of 0.01 x 10^(9) pN nm^2 for the elastic bending resistance of an individual microtubule, and 0.2 x 10^(9) pN nm^2 for the elastic bending resistance of the 9 + 2 region of the flagellum. An improvement in propulsive effectiveness by addition of a terminal filament remains to be demonstrated

Investigating the molecular mechanisms of organophosphate and pyrethroid resistance in the fall armyworm Spodoptera frugiperda.

Published onlineJournal ArticleResearch Support, Non-U.S. Gov'tThe fall armyworm Spodoptera frugiperda is an economically important pest of small grain crops that occurs in all maize growing regions of the Americas. The intensive use of chemical pesticides for its control has led to the selection of resistant populations, however, to date, the molecular mechanisms underlying resistance have not been characterised. In this study the mechanisms involved in the resistance of two S. frugiperda strains collected in Brazil to chlorpyrifos (OP strain) or lambda-cyhalothrin (PYR strain) were investigated using molecular and genomic approaches. To examine the possible role of target-site insensitivity the genes encoding the organophosphate (acetylcholinesterase, AChE) and pyrethroid (voltage-gated sodium channel, VGSC) target-site proteins were PCR amplified. Sequencing of the S. frugiperda ace-1 gene identified several nucleotide changes in the OP strain when compared to a susceptible reference strain (SUS). These result in three amino acid substitutions, A201S, G227A and F290V, that have all been shown previously to confer organophosphate resistance in several other insect species. Sequencing of the gene encoding the VGSC in the PYR strain, identified mutations that result in three amino acid substitutions, T929I, L932F and L1014F, all of which have been shown previously to confer knockdown/super knockdown-type resistance in several arthropod species. To investigate the possible role of metabolic detoxification in the resistant phenotype of the OP and PYR stains all EST sequences available for S. frugiperda were used to design a gene-expression microarray. This was then used to compare gene expression in the resistant strains with the susceptible reference strain. Members of several gene families, previously implicated in metabolic resistance in other insects were found to be overexpressed in the resistant strains including glutathione S-transferases, cytochrome P450s and carboxylesterases. Taken together these results provide evidence that both target-site and metabolic mechanisms underlie the resistance of S. frugiperda to pyrethroids and organophosphates.BBSRCNational Council for Scientific and Technological Development of Brazi

Larval dispersal of Spodoptera frugiperda strains on Bt Cotton: a model for understanding resistance evolution and consequences for its management.

High dispersal of Lepidoptera larvae between non-Bt and Bt cotton plants can favour the evolution of insect resistance; however, information on host acceptance of neonates in tropical transgenic crops is scarce. Therefore, the purposes of this study were as follows: (i) to investigate the feeding behaviour of susceptible and Cry1F-resistant strains of Spodoptera frugiperda (J.E. Smith) on Bt and non-Bt cotton (Gossypium hirsutum L.) varieties and (ii) to understand the possible effects of cotton field contamination on the dispersal and infestation capacity of S. frugiperda larvae by using an individualbased model. The main results of this paper are as follows: (1) the highest post-feeding larval dispersal of the Cry1F-resistant strain occurred at an exposure time of 18?24 h; (2) via video tracking assays, we found that the least distance moved was by larvae resistant to Cry1F on non-Bt cotton; and (3) the model indicated differences in mobility capacity between Bt and non-Bt cotton. We conclude that resistant neonates exhibit sedentary behaviour. Our report represents the first findings concerning the fitness cost of larval behaviour traits of S. frugiperda associated with Cry1F resistance in Brazilian populations.Publicado online em 23 de novembro de 2017

Genetic structure and molecular variability of Spodoptera frugiperda (Lepidoptera: Noctuidae) collected in maize and cotton fields in Brazil

The purpose of this research was to evaluate the genetic similarity and structure of the fall armyworm, Spodoptera frugiperda (J.E. Smith), populations associated with maize and cotton crops in Brazil using amplified fragment length polymorphisms. Mean genetic similarity among populations was 0.45. The unweighted pair group method with arithmetic mean analysis dendrograms did not separate populations of S. frugiperda into clusters related to the host plant in which the insects were collected. No genetic variation was observed among maize and cotton populations of S. frugiperda, suggesting that the same populations are injuring both crops in Brazil. This research validates the need for stewardship of crop-protection methods for managing S. frugiperda to reduce the incidence of pesticide resistance, due to the spatial and temporal overlapping of maize and cotton crops in some regions in Brazil

Analysis of Differential Efficacy and Affinity of GABAA (α1/α2) Selective Modulators.

Selective modulators of the γ-amino butyric acid (GABAA) family of receptors have the potential to treat a range of disease states related to cognition, pain, and anxiety. While the development of various α subunit-selective modulators is currently underway for the treatment of anxiety disorders, a mechanistic understanding of the correlation between their bioactivity and efficacy, based on ligand-target interactions, is currently still lacking. In order to alleviate this situation, in the current study we have analyzed, using ligand- and structure-based methods, a data set of 5440 GABAA modulators. The Spearman correlation (ρ) between binding activity and efficacy of compounds was calculated to be 0.008 and 0.31 against the α1 and α2 subunits of GABA receptor, respectively; in other words, the compounds had little diversity in structure and bioactivity, but they differed significantly in efficacy. Two compounds were selected as a case study for detailed interaction analysis due to the small difference in their structures and affinities (ΔpKi(comp1_α1 - comp2_α1) = 0.45 log units, ΔpKi(comp1_α2 - comp2_α2) = 0 log units) as compared to larger relative efficacies (ΔRE(comp1_α1 - comp2_α1) = 1.03, ΔRE(comp1_α2 - comp2_α2) = 0.21). Docking analysis suggested that His-101 is involved in a characteristic interaction of the α1 receptor with both compounds 1 and 2. Residues such as Phe-77, Thr-142, Asn-60, and Arg-144 of the γ chain of the α1γ2 complex also showed interactions with heterocyclic rings of both compounds 1 and 2, but these interactions were disturbed in the case of α2γ2 complex docking results. Binding pocket stability analysis based on molecular dynamics identified three substitutions in the loop C region of the α2 subunit, namely, G200E, I201T, and V202I, causing a reduction in the flexibility of α2 compared to α1. These amino acids in α2, as compared to α1, were also observed to decrease the vibrational and dihedral entropy and to increase the hydrogen bond content in α2 in the apo state. However, freezing of both α1 and α2 was observed in the ligand-bound state, with an increased number of internal hydrogen bonds and increased entropy. Therefore, we hypothesize that the amino acid differences in the loop C region of α2 are responsible for conformational changes in the protein structure compared to α1, as well as for the binding modes of compounds and hence their functional signaling

The optimal elastic flagellum

Motile eukaryotic cells propel themselves in viscous fluids by passing waves of bending deformation down their flagella. An infinitely long flagellum achieves a hydrodynamically optimal low-Reynolds number locomotion when the angle between its local tangent and the swimming direction remains constant along its length. Optimal flagella therefore adopt the shape of a helix in three dimensions (smooth) and that of a sawtooth in two dimensions (non-smooth). Physically, biological organisms (or engineered micro-swimmers) must expend internal energy in order to produce the waves of deformation responsible for the motion. Here we propose a physically-motivated derivation of the optimal flagellum shape. We determine analytically and numerically the shape of the flagellar wave which leads to the fastest swimming while minimizing an appropriately-defined energetic expenditure. Our novel approach is to define an energy which includes not only the work against the surrounding fluid, but also (1) the energy stored elastically in the bending of the flagellum, (2) the energy stored elastically in the internal sliding of the polymeric filaments which are responsible for the generation of the bending waves (microtubules), and (3) the viscous dissipation due to the presence of an internal fluid. This approach regularizes the optimal sawtooth shape for two-dimensional deformation at the expense of a small loss in hydrodynamic efficiency. The optimal waveforms of finite-size flagella are shown to depend upon a competition between rotational motions and bending costs, and we observe a surprising bias towards half-integer wave-numbers. Their final hydrodynamic efficiencies are above 6%, significantly larger than those of swimming cells, therefore indicating available room for further biological tuning

The frequency in Japanese of genetic variants of 22 proteins III. Phosphoglucomutase-1, phosphoglucomutase-2, 6-phosphogluconate dehydrogenase, adenylate kinase, and adenosine deaminase

Five enzyme systems, PGM 1 , PGM 2 , ADA, 6-PGD and AK, were examined by electrophoresis in over 4000 samples from Hiroshima and Nagasaki for the frequencies of common and rare variants. In the PGM 1 , system, the PGM 2 1 allele and PGM 7 1 ; allele were found in polymorphic proportions. I n addition, five kinds of slow variants and three types of fast variants of PGM 1 were detected. The PGM 3 NGS 1 1 allele was found in five individuals from Nagasaki, but was not observed in samples from Hiroshima. There were no variants of PGM 2 . Three kinds of fast variants of 6-PGD were detected. NO variation in AK was observed. There were no rare variants of ADA. The 6-PGD c allele had a frequency of 0.084 in Hiroshima, and 0.093 in Nagasaki, and the ADA 2 allele frequencies of 0.025 in Hiroshima and 0.032 in Nagasaki.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65524/1/j.1469-1809.1977.tb01912.x.pd

Manejo da resistência de Bonagota salubricola e Grapholita a inseticidas.

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Metodologias para instalação de bioensaios para o monitoramento da resistência de bonagota salubricola e grapholita molesta a inseticidas.

Neste comunicado técnico, são apresentadas metodologias de bioensaios que podem ser empregadas para mensurar a suscetibilidade de Bonagota salubricola e Grapholita molesta a inseticidas, visando a disponibilizar técnicas para implementar um programa pró-ativo de manejo da resistência.bitstream/item/104060/1/Comunicado-Tecnico-152.pd

Exploitation of mitochondrial nad6 as a complementary marker for studying population variability in Lepidoptera

The applicability of mitochondrial nad6 sequences to studies of DNA and population variability in Lepidoptera was tested in four species of economically important moths and one of wild butterflies. The genetic information so obtained was compared to that of cox1 sequences for two species of Lepidoptera. nad6 primers appropriately amplified all the tested DNA targets, the generated data proving to be as informative and suitable in recovering population structures as that of cox1. The proposal is that, to obtain more robust results, this mitochondrial region can be complementarily used with other molecular sequences in studies of low level phylogeny and population genetics in Lepidoptera