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Solid-phase microextraction liquid chromatography/tandem mass spectrometry to determine postharvest fungicides in fruits.

A method to determine five postharvest fungicides (dichloran, flutriafol, o-phenylphenol, prochloraz, tolclofos methyl) in fruits (cherries, lemons, oranges, peaches) has been developed using solid-phase microextraction (SPME) coupled to liquid chromatography (LC) with photodiode array (DAD), mass spectrometry (MS), or tandem mass spectrometry (MS/MS) with ion trap detection. Extraction involved sample homogenization with an acetone/water solution (5:1), filtration, and acetone evaporation prior to fiber extraction. The pesticides were isolated with a fused-silica fiber coated with 50-microm Carbowax/template resin. The effects of pH, ion strength, sample volume, and extraction time were investigated, and their impact on the SPME-LC/MS was studied. Dynamic and static modes of desorption were compared and the variables affecting desorption processes in SPME-LC optimized. Static desorption provided the best recoveries and peak shapes. Recoveries at the limit of quantification (LOQ) levels were between 10% for prochloraz and 60% for o-phenylphenol, with relative standard deviations from 13.6% for prochloraz to 3.1% for o-phenylphenol. The versatility of the method was also exhibited by its excellent linearity in the concentration intervals between 0.0005 and 5 mg kg(-1) for dichloran and 0.01-10 mg kg(-1) for tolclofos methyl and prochloraz. LOQs ranged from 0.25 to 1 microg g(-1) using DAD, from 0.002 to 0.01 microg g(-1) using LC/MS, and from 0.0005 to 0.01 to microg g(-1) using LC/MS/MS. LOQs obtained in the present study using LC/MS and LC/MS/MS are lower than maximum residue limits established for all the fungicides in any matrix studied. The method enables to determine polar pesticides at low-microgram per gram levels in fruits.

Calibration↗

It takes guts: the Drosophila hindgut as a model system for organogenesis.

The Drosophila hindgut is fruitful territory for investigation of events common to many types of organogenesis. The development of the Drosophila hindgut provides, in microcosm, a genetic model system for studying processes such as establishment (patterning) of an epithelial primordium, its internalization by gastrulation, development of left--right asymmetric looping, patterning in both the anteroposterior and dorsoventral axes, innervation, investment of an epithelium with mesoderm, reciprocal epitheliomesenchymal interactions, cell shape change, and cell rearrangement. We review the genetic control of these processes during development of the Drosophila hindgut, and compare these to related processes in other bilaterians, particularly vertebrates. We propose that caudal/Cdx, brachyenteron/Brachyury, fork head/HNF-3, and wingless/Wnt constitute a conserved "cassette" of genes expressed in the blastopore and later in the gut, involved in posterior patterning, cell rearrangement, and gut maintenance. Elongation of the internalized Drosophila hindgut primordium is similar to elongation of the archenteron and also of the entire embryonic axis (both during and after gastrulation), as well as of various tubules (e.g., nephric ducts, Malpighian tubules), as it is driven by cell rearrangement. The genes drumstick, bowl, and lines (which encode putative transcriptional regulators) are required for this cell rearrangement, as well as for spatially localized gene expression required to establish the three morphologically distinct subregions of the hindgut. Expression of signaling molecules regulated by drumstick, bowl, and lines, in particular of the JAK/STAT activator Unpaired at the hindgut anterior, may play a role in controlling hindgut cell rearrangement. Other cell signaling molecules expressed in the hindgut epithelium are required to establish its normal size (Dpp and Hh), and to establish and maintain the hindgut visceral mesoderm (Wg and Hh). Both maternal gene activity and zygotic gene activity are required for asymmetric left--right looping of the hindgut. Some of the same genes (caudal and brachyenteron) required for embryonic hindgut development also act during pupation to construct a new hindgut from imaginal cells. Application of the plethora of genetic techniques available in Drosophila, including forward genetic screens, should identify additional genes controlling hindgut development and thus shed light on a variety of common morphogenetic processes.

Animals↗

Direct Access to Plant Epicuticular Wax Crystals by a New Mechanical Isolation Method.

A new method for the isolation of wax crystals from plant surfaces is presented. The wax-covered plant surface, e.g., a piece of a leaf or fruit, is brought into contact with a preparation liquid, e.g., glycerol or triethylene glycol, and cooled to ca. -100 degrees C. When the plant specimen is removed, the epicuticular wax remains embedded in the frozen liquid. After it warms up, the wax layer can be captured on appropriate carriers for further studies. This isolation method causes very little stress on the wax crystals; thus the shape and crystal structure are well preserved. In many cases it is possible, by choosing a preparation liquid with appropriate wettability, to isolate either the entire epicuticular wax layer or only discrete wax crystals without the underlying wax film. These crystals are well suited for electron diffraction studies by transmission electron microscopy and high resolution imaging by atomic force microscopy. The absence of intracuticular components and other impurities and the feasibility of the selective isolation of wax crystals enable improved chemical analysis and a more detailed study of their properties.

Journal Article↗

Rho GTPase function in flies: insights from a developmental and organismal perspective.

Morphogenesis is a key event in the development of a multicellular organism and is reliant on coordinated transcriptional and signal transduction events. To establish the segmented body plan that underlies much of metazoan development, individual cells and groups of cells must respond to exogenous signals with complex movements and shape changes. One class of proteins that plays a pivotal role in the interpretation of extracellular cues into cellular behavior is the Rho family of small GTPases. These molecular switches are essential components of a growing number of signaling pathways, many of which regulate actin cytoskeletal remodeling. Much of our understanding of Rho biology has come from work done in cell culture. More recently, the fruit fly Drosophila melanogaster has emerged as an excellent genetic system for the study of these proteins in a developmental and organismal context. Studies in flies have greatly enhanced our understanding of pathways involving Rho GTPases and their roles in development.

Animals↗

[The computer image processing and cluster analysis on Fructus Cnidii from different habitats].

OBJECTIVE: To study the variation of shape of Fructus Cnidii from different habitats. METHODS: The methods of computer image processing and cluster analysis were adopted. RESULTS: The shape of Fructus Cnidii from different habitats varied significantly. CONCLUSION: According to the results of computer image processing and cluster analysis, Fructus Cnidii can be classified into three types.

China↗

Steroid-dependent modification of Hox function drives myocyte reprogramming in the Drosophila heart.

In the Drosophila larval cardiac tube, aorta and heart differentiation are controlled by the Hox genes Ultrabithorax (Ubx) and abdominal A (abdA), respectively. There is evidence that the cardiac tube undergoes extensive morphological and functional changes during metamorphosis to form the adult organ, but both the origin of adult cardiac tube myocytes and the underlying genetic control have not been established. Using in vivo time-lapse analysis, we show that the adult fruit fly cardiac tube is formed during metamorphosis by the reprogramming of differentiated and already functional larval cardiomyocytes, without cell proliferation. We characterise the genetic control of the process, which is cell autonomously ensured by the modulation of Ubx expression and AbdA activity. Larval aorta myocytes are remodelled to differentiate into the functional adult heart, in a process that requires the regulation of Ubx expression. Conversely, the shape, polarity, function and molecular characteristics of the surviving larval contractile heart myocytes are profoundly transformed as these cells are reprogrammed to form the adult terminal chamber. This process is mediated by the regulation of AbdA protein function, which is successively required within these persisting myocytes for the acquisition of both larval and adult differentiated states. Importantly, AbdA specificity is switched at metamorphosis to induce a novel genetic program that leads to differentiation of the terminal chamber. Finally, the steroid hormone ecdysone controls cardiac tube remodelling by impinging on both the regulation of Ubx expression and the modification of AbdA function. Our results shed light on the genetic control of one in vivo occurring remodelling process, which involves a steroid-dependent modification of Hox expression and function.

Animals↗

Endocrine disruptors and sexually dimorphic behaviors: a question of heads and tails.

Early brain development seems destined to be the territory where questions about the neurotoxicity of endocrine disruptors will be contested, so we ought to clarify how we propose to invade it. Everything else we have learned tells us that the most sensitive indices are likely to be the most subtle, a term we often use to describe the phenomena and domain of neurobehavioral toxicology. It might mean an effect that is small in amplitude, or that requires sensitive measures to detect. One class of effects that might result from environmental exposure to endocrine disruptors reflects both kinds of subtlety. It consists of sexually dimorphic behaviors that seem biologically driven but that could easily masquerade as socially-defined or gender differences. Performances on neuropsychological tests fall into this category. Beyond debates about their origins, such differences tend to be relatively modest in size, especially by the routine criteria adopted by toxicologists. Overwhelmingly, they rely on differences in means accompanied by p values. Such criteria are inappropriate for assessing hypotheses or calculating risks based on the impact of what invariably will be quite low environmental levels. In this arena at least, the quantification of effects should proceed from examinations of distributions. Instead of conventional descriptions based on measures of central tendency, we should instead examine the shapes of distributions, especially the tails. Neuropsychological differences between males and females are amplified and emerge at the extremes, even when sex accounts for no more than a minuscule proportion of the variance. Lead taught a similar lesson about how to quantify population impacts. Furthermore, such differences at the extremes suggest both practical and theoretical implications that can too easily be ignored but that provide paths for fruitful exploration.

Animals↗

The fruits of ill-health: Pesticides and workers' bodies in post-World War II California.

In the postwar period, modernist frameworks of the human body, which described the body as both cosmopolitan and separated from its environment, competed with ecological frameworks that constructed the body as inherently porous and tightly linked to the surrounding world. The history of pesticide-related illness among farmworkers, and the gradual recognition that pesticides posed a new kind of public health problem, illustrates how these competing understandings were adopted, mobilized, and applied by different groups, as well as how politics shaped the emergence of new medical facts. New forms of illness generated new knowledge about the modern landscape and made visible material links between bodies and their environments.

Agricultural Workers' Diseases↗

Dental microwear in anubis and hybrid baboons (Papio hamadryas, sensu lato) living in Awash National Park, Ethiopia.

We describe dental microwear in baboons (Papio hamadryas sensu lato) from the anubis-hamadryas hybrid zone of Awash National Park, Ethiopia, outline its variation with sex and age, and attempt to relate the observed microwear pattern to environment and diet. Casts of the maxillary second molar of 52 adult and subadult individuals of both sexes were examined with a scanning electron microscope at x 500. Digitized micrographs were taken at a consistent location on facet 9, and microwear was recorded with an image analysis software package. Univariate and multivariate statistics were used to investigate the shape, size, and density of microwear features. The overall pattern of microwear exhibits an unusual combination of high feature density, with numerous small pits and relatively wide striations, and a high correlation between width of pits and striations across individuals. We interpret this pattern as predominantly the consequence of abrasion by relatively small-caliber environmental grit when accidentally ingested with tough foods such as dried seeds and fruits, as expected in a terrestrial omnivore living in a dusty habitat. Statistical analysis revealed no significant differences between groups defined by sex, age, or troop membership, a result consistent with qualitative observations of feeding habits in this population, and which lends no support to the hypothesis that the longer jaws of adult males should result in longer striations. A trend towards greater feature density in females, however, might be due to limited sexual dinichism, and merits further investigation.

Analysis of Variance↗

Structure and morphogenesis of the eggshell and micropylar apparatus in the olive fly, Dacus oleae (Diptera: Tephritidae).

The egg of the olive fly, Dacus oleae (Diptera, Tephritidae), is laid inside olives and the larva eventually destroys the fruit. The oocyte is surrounded by several distinct layers which are produced during choriogenesis. The chorion covering the main body of the egg outside of the vitelline membrane includes a "wax" layer, an innermost chorionic layer, an endochorion consisting of inner and outer layers separated by pillars and cavities similar to their counterparts in Drosophila melanogaster, as well as inner and outer exochorionic layers. The anterior pole is shaped like an inverted cup, which is chiefly hollow around its base and has very large openings communicating with the environment. Holes through the surface of the endochorion result from deposition of endochorionic substance around follicular cell microvilli. An opening at the apex of the cup provides an entrance for sperm entering the micropylar canal, which traverses the endochorion and continues into a "pocket" in a thickened vitelline protrusion. The micropylar canal is formed by deposition of endochorion and vitelline membrane around an elongated pair of follicular cell extensions. These extensions later degenerate and leave an empty canal about 5 microns in diameter and the narrower pocket about 1 micron in diameter. Respiration is thought to be facilitated by openings at the base of the anterior pole as well as by openings through the "plastron" around the main body of the shell.

Animals↗

Maternal and developmental temperature modulate adult response to temperature in Drosophila melanogaster.

Beyond inherited genes and environmentally induced changes in gene expression, phenotypes can also be shaped by parental effects-an effect from a parental phenotype that causes modifications in offspring traits, which cannot be solely explained by the parental or offspring genomes. Such effects may prepare offspring for future environmental conditions and contribute to phenotypic plasticity, including responses to temperature. While temperature-induced plasticity has been extensively studied, the relative contributions of parental versus direct environmental cues remain poorly understood. The fruit fly Drosophila melanogaster is a powerful model for studying physiological and behavioral adaptation to temperature. Flies inhabit environments spanning broad thermal ranges and show evidence of parental effects, such as increased heat tolerance in offspring from warm-reared parents. Here, we exposed mothers to two experimental temperatures and split their broods between the same two temperatures to estimate the relative importance of maternal and developmental effects on adult physiological and developmental responses to temperature. We find that the reaction norms of locomotor activity under gradually increasing temperatures, responses to heat-shock and cold-shock, and fecundity are mostly governed by direct plastic responses to developmental environment. We detected comparatively weak maternal effects in the response to heat-shock, fecundity, and grand-offspring survival where matched environments counteracted the effects of direct offspring experience. We conclude that thermal experience during development is the primary determinant of phenotypic plasticity in D. melanogaster, while maternal experience contributes a small but non-negligible component.

Animals↗

A prokaryotic dnaA sequence in Drosophila melanogaster: Wolbachia infection and cytoplasmic incompatibility among laboratory strains.

Using oligonucleotide primers derived from the aligned polypeptide sequences of several prokaryotic dnaA genes, we amplified from Drosophila melanogaster DNA a 557 bp fragment containing a single open reading frame. The predicted peptide sequence shows a significant similarity to previously characterized protein sequences that are encoded by the dnaA genes of several prokaryotes. The dnaA sequences are also detectable by PCR in DNA from Drosophila simulans and Nasonia vitripennis flies which are infected by a symbiotic bacterium assigned to the type species Wolbachia pipientis. A tetracycline treatment that eradicates bacterial parasites from insects, abolishes the dnaA sequences from Drosophila and Nasonia DNA. In addition, dnaA-positive Drosophila melanogaster contain numerous rod-shaped bacteria in embryos, which are abolished in subsequent generations after treatment with tetracycline. Combined with phylogenetic analysis of DnaA and 16S rRNA sequences, these results show that the dnaA cognate comes from Wolbachia. A survey of Drosophila stocks using PCR amplification of dnaA and 16S rRNA sequences showed that Wolbachia is widely spread among D. melanogaster laboratory strains but absent from several established strains of the Mediterranean fruit fly Ceratitis capitata. Evidence is also presented that presence of the bacterium can cause partial cytoplasmic incompatibility between infected and non-infected D. melanogaster strains.

Amino Acid Sequence↗

[The differences and regularities of life-span dynamics for human and animal populations].

The dynamics of mortality of several generations of man, Wistar rats and line Canton-S fruit flies Drosophila melanogaster was investigated. The result was, that the correlation between sample mean and sample variance was negative for man, positive for flies and was absent for rats. To fit the survival curve we chose two-parametric Gompertz function. Starting from Strehler-Mildwan correlation (linear correlation between Gompertz function parameters), we obtained theoretical dependence between life-span mean and variance. This dependence had complex shape and points, corresponding to different species were situated at different segments of this curve that is corresponding to the various sign of correlation between these characteristics. Increasing of the mean man life-span attribute usually to advances in medicine and social environment. Associated with this the decreasing of the life-span variance, as it follows from this paper, depends on parameter point location on parametric plane, i.e. from biological feature of species.

Animals↗

Reproductive biology and genetic diversity of a cryptoviviparous mangrove aegiceras corniculatum (Myrsinaceae) using allozyme and intersimple sequence repeat (ISSR) analysis

Mangroves consist of a group of taxonomically diverse species representing about 20 families of angiosperms. However, little is known about their reproductive biology, genetic structure, and the ecological and genetic factors affecting this structure. Comparative studies of various mangrove species are needed to fill such gaps in our knowledge. The pollination biology, outcrossing rate, and genetic diversity of Aegiceras corniculatum were investigated in this study. Pollination experiments suggested that the species is predominantly pollinator-dependent in fruit setting. A quantitative analysis of the mating system was performed using progeny arrays assayed for intersimple sequence repeat (ISSR) markers. The multilocus outcrossing rate (tm) was estimated to be 0.653 in a wild population. Both allozyme and ISSR were used to investigate genetic variation within and among populations. The combined effects of founder events and enhanced local gene flow through seedling dispersal by ocean currents apparently played an important role in shaping the population genetic structure in this mangrove species. Both allozyme variation (P = 4.76%, A = 1.05, HE = 0.024) and ISSR diversity (P = 16.18%, A = 1.061, HE = 0.039) were very low at the species level, in comparison with other woody plants with mixed-mating or outcrossing systems. Gene differentiation among populations was also low: allozyme GST = 0.106 and ISSR GST = 0.178. The unusually high genetic identities (0.997 for allozyme and 0.992 for ISSR loci), however, suggest that these populations are probably all descended from a common ancestral population with low polymorphism.

Journal Article↗

Morphometrics and the role of the phenotype in studies of the evolution of developmental mechanisms.

Developmental mechanisms are usually assumed to evolve by natural selection of the morphological traits they produce. Therefore, information on phenotypic traits is an important component of comparative studies of development. Morphometrics permits the rigorous quantitative analysis of variation in organismal size and shape, and is increasingly being used in developmental contexts. The new methods of morphometrics combine a geometric concept of shape with the procedures of multivariate statistics, and constitute a powerful and flexible set of tools for analyzing morphological variation. This paper briefly reviews these methods and provides examples of their application in studies of genetic variation and developmental modularity. The results of morphometric analyses can be readily interpreted in relation to the geometry and anatomical structure of the parts under study. Genetic studies of shape in the mouse mandible found two recurrent patterns in environmental and genetic variation from different origins, suggesting that the development system 'channels' the phenotypic expression of variation in similar ways. Moreover, by analyzing the correlations of left-right asymmetries of morphometric traits, it is possible to delimit the spatial extent of developmental modules. These methods complement the experimental approaches of developmental biology and genetics, and can be expected to be especially fruitful in combination with them.

Animals↗

Testing for genetic evidence of population expansion and contraction: an empirical analysis of microsatellite DNA variation using a hierarchical Bayesian model.

The role of past climatic change in shaping the distributions of tropical rain forest vertebrates is central to long-standing hypotheses about the legacy of the Quaternary ice ages. One approach to testing such hypotheses is to use genetic data to infer the demographic history of codistributed species. Population genetic theory that relates the structure of allelic genealogies to historical changes in effective population size can be used to detect a past history of demographic expansion or contraction. The fruit bats Cynopterus sphinx and C. brachyotis (Chiroptera: Pteropodidae) exhibit markedly different distribution patterns across the Indomalayan region and therefore represent an exemplary species pair to use for such tests. The purpose of this study was to test alternative hypotheses about historical patterns of demographic expansion and contraction in C. sphinx and C. brachyotis using a coalescent-based analysis of microsatellite variation. Specifically, we used a hierarchical Bayesian model based on Markov chain Monte Carlo simulations to estimate the posterior distribution of genealogical and demographic parameters. The results revealed strong evidence for population contraction in both species. Evidence for a population contraction in C. brachyotis was expected on the basis of biogeographic considerations. However, similar evidence for population contraction in C. sphinx does not support the hypothesis that this species underwent a pronounced range expansion during the late Quaternary. Genetic evidence for population decline may reflect the consequences of habitat destruction on a more recent time scale.

Animals↗

Reexamination of the role of autolysis in the development of Myxococcus xanthus.

It has been widely reported that 80 to 90% of the cell population undergoes autolysis during sporulation in Myxococcus xanthus. A re-evaluation of the techniques used to measure autolysis in M. xanthus showed that the methods previously used to draw this conclusion are subject to artifacts, which result in a substantial underestimation of the number of cells present during development. We found that at least 80% of the cells that enter development survive throughout fruiting body formation. The cell loss that did occur appeared to be gradual over a period of at least 7 days. Our results suggest that autolysis is not an obligate stage in the development of M. xanthus. The data also showed that sporulating cells pass through a prespore stage in which they become osmotically and physically fragile and therefore difficult to harvest intact. The fragility was correlated with the change from a rod to a spherical shape. As the prespores differentiated into refractile spores, they lost fragility and became amenable to harvesting by standard protocols.

Bacteriolysis↗