Showing papers by "Washington State University published in 2006"

Reconstructing the early evolution of Fungi using a six-gene phylogeny

Abstract: The ancestors of fungi are believed to be simple aquatic forms with flagellated spores, similar to members of the extant phylum Chytridiomycota (chytrids). Current classifications assume that chytrids form an early-diverging clade within the kingdom Fungi and imply a single loss of the spore flagellum, leading to the diversification of terrestrial fungi. Here we develop phylogenetic hypotheses for Fungi using data from six gene regions and nearly 200 species. Our results indicate that there may have been at least four independent losses of the flagellum in the kingdom Fungi. These losses of swimming spores coincided with the evolution of new mechanisms of spore dispersal, such as aerial dispersal in mycelial groups and polar tube eversion in the microsporidia (unicellular forms that lack mitochondria). The enigmatic microsporidia seem to be derived from an endoparasitic chytrid ancestor similar to Rozella allomycis, on the earliest diverging branch of the fungal phylogenetic tree.

Insights into social insects from the genome of the honeybee Apis mellifera

Abstract: Here we report the genome sequence of the honeybee Apis mellifera, a key model for social behaviour and essential to global ecology through pollination. Compared with other sequenced insect genomes, the A. mellifera genome has high A+T and CpG contents, lacks major transposon families, evolves more slowly, and is more similar to vertebrates for circadian rhythm, RNA interference and DNA methylation genes, among others. Furthermore, A. mellifera has fewer genes for innate immunity, detoxification enzymes, cuticle-forming proteins and gustatory receptors, more genes for odorant receptors, and novel genes for nectar and pollen utilization, consistent with its ecology and social organization. Compared to Drosophila, genes in early developmental pathways differ in Apis, whereas similarities exist for functions that differ markedly, such as sex determination, brain function and behaviour. Population genetics suggests a novel African origin for the species A. mellifera and insights into whether Africanized bees spread throughout the New World via hybridization or displacement.

Biological invasions: recommendations for U.S. policy and management.

Abstract: The Ecological Society of America has evaluated current U.S. national policies and practices on biological invasions in light of current scientific knowledge. Invasions by harmful nonnative species are increasing in number and area affected; the damages to ecosystems, economic activity, and human welfare are accumulating. Without improved strategies based on recent scientific advances and increased investments to counter invasions, harm from invasive species is likely to accelerate. Federal leadership, with the cooperation of state and local governments, is required to increase the effectiveness of prevention of invasions, detect and respond quickly to new potentially harmful invasions, control and slow the spread of existing invasions, and provide a national center to ensure that these efforts are coordinated and cost effective. Specifically, the Ecological Society of America recommends that the federal government take the following six actions: (1) Use new information and practices to better manage commercial and other pathways to reduce the transport and release of potentially harmful species; (2) Adopt more quantitative procedures for risk analysis and apply them to every species proposed for importation into the country; (3) Use new cost-effective diagnostic technologies to increase active surveillance and sharing of information about invasive species so that responses to new invasions can be more rapid and effective; (4) Create new legal authority and provide emergency funding to support rapid responses to emerging invasions; (5) Provide funding and incentives for cost-effective programs to slow the spread of existing invasive species in order to protect still uninvaded ecosystems, social and industrial infrastructure, and human welfare; and (6) Establish a National Center for Invasive Species Management (under the existing National Invasive Species Council) to coordinate and lead improvements in federal, state, and international policies on invasive species. Recent scientific and technical advances provide a sound basis for more cost-effective national responses to invasive species. Greater investments in improved technology and management practices would be more than repaid by reduced damages from current and future invasive species. The Ecological Society of America is committed to assist all levels of government and provide scientific advice to improve all aspects of invasive-species management.

Continuities and Extensions of Ethical Climate Theory: A Meta-Analytic Review

Abstract: Using traditional meta-analytic techniques, we compile relevant research to enhance conceptual appreciation of ethical climate theory (ECT) as it has been studied in the descriptive and applied ethics literature. We explore the various treatments of ethical climate to understand how the theoretical framework has developed. Furthermore, we provide a comprehensive picture of how the theory has been extended by describing the individual-level work climate outcomes commonly studied in this theoretical context. Meta-analysis allows us to resolve inconsistencies in previous findings as well as confirm the central tenets of the overall ethical climate framework. In addition, we consider the ethical climate relationships in the larger context of the␣theoretical framework, using path analysis to test the structural relationships. Overall, our results provide evidence of the relationships between ethical climate perceptions and individual-level work outcomes. Based on our analyses, we offer future research directions important for further development of ECT.

Trends in microwave- related drying of fruits and vegetables

Abstract: Microwave (MW)-related (MW-assisted or MW-enhanced) combination drying is a rapid dehydration technique that can be applied to specific foods, particularly to fruits and vegetables. Increasing concerns over product quality and production costs have motivated the researchers to investigate and the industry to adopt combination drying technologies. The advantages of MW-related combination drying include the following: shorter drying time, improved product quality, and flexibility in producing a wide variety of dried products. But current applications are limited to small categories of fruits and vegetables due to high start-up costs and relatively complicated technology as compared to conventional convection drying. MW-related combination drying takes advantages of conventional drying methods and microwave heating, leading to better processes than MW drying alone. This paper presents a comprehensive review of recent progresses in MW-related combined drying research and recommendations for future research to bridge the gap between laboratory research and industrial applications.

On the spectroscopic and thermochemical properties of ClO, BrO, IO, and their anions.

Abstract: A coupled cluster composite approach has been used to accurately determine the spectroscopic constants, bond dissociation energies, and heats of formation for the X12Π3/2 states of the halogen oxides ClO, BrO, and IO, as well as their negative ions ClO-, BrO-, and IO-. After determining the frozen core, complete basis set (CBS) limit CCSD(T) values, corrections were added for core−valence correlation, relativistic effects (scalar and spin−orbit), the pseudopotential approximation (BrO and IO), iterative connected triple excitations (CCSDT), and iterative quadruples (CCSDTQ). The final ab initio equilibrium bond lengths and harmonic frequencies for ClO and BrO differ from their accurate experimental values by an average of just 0.0005 A and 0.8 cm-1, respectively. The bond length of IO is overestimated by 0.0047 A, presumably due to an underestimation of molecular spin−orbit coupling effects. Spectroscopic constants for the spin−orbit excited X22Π1/2 states are also reported for each species. The predicted...

Academic Dishonesty in Graduate Business Programs: Prevalence, Causes, and Proposed Action

Abstract: Little is currently known about cheating among graduate business students. We collected data from more than 5,000 business (mostly MBA) and nonbusiness graduate students at 32 colleges and universities in the United States and Canada during the 2002–2003 and 2003–2004 academic years to test a series of hypotheses regarding the prevalence of graduate business student cheating and reasons why these students cheat. We found that graduate business students cheat more than their nonbusiness-student peers. Correlation results found cheating to be associated with perceived peer behavior, as well as the perceived certainty of being reported by a peer, and the understanding and acceptance of academic integrity policies by students and faculty. But, regression analysis results suggest that perceived peer behavior has the largest effect. Drawing from these findings and past research on undergraduate students, we propose strategies that business schools and faculty can use to promote academic integrity in graduate business programs.

The ACS Virgo Cluster survey. VI. Isophotal analysis and the structure of early-type galaxies

Abstract: We present a detailed analysis of the morphology, isophotal parameters, and surface brightness profiles for 100 early-type members of the Virgo Cluster, from dwarfs (MB = -15.1 mag) to giants (MB = -21.8 mag), imaged in the g and z passbands using the Advanced Camera for Surveys on board the Hubble Space Telescope. Dust and complex morphological structures are common. Dust is detected in 42% of galaxies brighter than BT = 12.15 mag, while kiloparsec-scale stellar disk, bars, and nuclear stellar disks are seen in 60% of galaxies with intermediate luminosity. Isophotal parameters are derived typically within 8 kpc from the center for the brightest galaxies, and 1.5 kpc for the faintest systems, with a resolution of 7 pc. For most galaxies, the surface brightness profiles are well described by a Sersic model with index n that increases steadily with the galaxy luminosity; only for 8 of the 10 brightest galaxies are the inner profiles (typically within 100 pc of the center) lower than expected based on an extrapolation of the outer Sersic model, and are better described by a single power-law function. Contrary to previous claims, we find no evidence in support of a strong bimodal behavior of the logarithmic slope of the inner surface brightness profile, γ; in particular the γ distribution for galaxies that do not show evidence of multiple morphological components is unimodal across the entire magnitude range spanned by the ACSVCS galaxies. Although the brightest galaxies have shallow inner profiles, the shallowest profiles are found in faint dwarf systems. The widely adopted separation of early-type galaxies between core and power-law types is questioned based on the present study.

The ACS Virgo Cluster Survey. VIII. The Nuclei of Early-Type Galaxies*

Abstract: The ACS Virgo Cluster Survey is a Hubble Space Telescope program to obtain high-resolution imaging in widely separated bandpasses (F475W ≈ g and F850LP ≈ z) for 100 early-type members of the Virgo Cluster, spanning a range of 460 in blue luminosity. We use this large, homogenous data set to examine the innermost structure of these galaxies and to characterize the properties of their compact central nuclei. We present a sharp upward revision in the frequency of nucleation in early-type galaxies brighter than M_B ≈ -15 (66% ≾ f_n ≾ 82%) and show that ground-based surveys underestimated the number of nuclei due to surface brightness selection effects, limited sensitivity and poor spatial resolution. We speculate that previously reported claims that nucleated dwarfs are more concentrated toward the center of Virgo than their nonnucleated counterparts may be an artifact of these selection effects. There is no clear evidence from the properties of the nuclei, or from the overall incidence of nucleation, for a change at M_B ~ -17.6, the traditional dividing point between dwarf and giant galaxies. There does, however, appear to be a fundamental transition at M_B ~ -20.5, in the sense that the brighter, "core-Sersic" galaxies lack resolved (stellar) nuclei. A search for nuclei that may be offset from the photocenters of their host galaxies reveals only five candidates with displacements of more than 0"5, all of which are in dwarf galaxies. In each case, however, the evidence suggests that these "nuclei" are, in fact, globular clusters projected close to the galaxy photocenter. Working from a sample of 51 galaxies with prominent nuclei, we find a median half-light radius of (r_h) = 4.2 pc, with the sizes of individual nuclei ranging from 62 pc down to ≤2 pc (i.e., unresolved in our images) in about a half-dozen cases. Excluding these unresolved objects, the nuclei sizes are found to depend on nuclear luminosity according to the relation r_h α L ^(0.50±0.03). Because the large majority of nuclei are resolved, we can rule out low-level AGNs as an explanation for the central luminosity excess in almost all cases. On average, the nuclei are ≈3.5 mag brighter than a typical globular cluster. Based on their broadband colors, the nuclei appear to have old to intermediate age stellar populations. The colors of the nuclei in galaxies fainter than M_B ≈ -17.6 are tightly correlated with their luminosities, and less so with the luminosities of their host galaxies, suggesting that their chemical enrichment histories were governed by local or internal factors. Comparing the nuclei to the "nuclear clusters" found in late-type spiral galaxies reveals a close match in terms of size, luminosity, and overall frequency. A formation mechanism that is rather insensitive to the detailed properties of the host galaxy properties is required to explain this ubiquity and homogeneity. The mean of the frequency function for the nucleus-to-galaxy luminosity ratio in our nucleated galaxies, (log η) = -2.49 ± 0.09 dex (σ = 0.59 ± 0.10), is indistinguishable from that of the SBH-to-bulge mass ratio, log(M/M_(gal) = -2.61 ± 0.07 dex (σ = 0.45 ± 0.09), calculated in 23 early-type galaxies with detected supermassive black holes (SBHs). We argue that the compact stellar nuclei found in many of our program galaxies are the low-mass counterparts of the SBHs detected in the bright galaxies. If this interpretation is correct, then one should think in terms of "central massive objects"—either SBHs or compact stellar nuclei—that accompany the formation of almost all early-type galaxies and contain a mean fraction ≈0.3% of the total bulge mass. In this view, SBHs would be the dominant formation mode above M_B ≈ -20.5.

An endogenous peptide signal in Arabidopsis activates components of the innate immune response.

Abstract: Innate immunity is initiated in animals and plants through the recognition of a variety of pathogen-associated molecules that in animals are called pathogen-associated molecular patterns and in plants are called elicitors. Some plant pathogen-derived elicitors have been identified as peptides, but peptide elicitors derived from the plant itself that activate defensive genes against pathogens have not been previously identified. Here, we report the isolation and characterization of a 23-aa peptide from Arabidopsis, called AtPep1, which activates transcription of the defensive gene defensin (PDF1.2) and activates the synthesis of H2O2, both being components of the innate immune response. The peptide is derived from a 92-aa precursor encoded within a small gene that is inducible by wounding, methyl jasmonate, and ethylene. Constitutive expression of the AtPep1 precursor gene PROPEP1 in transgenic Arabidopsis plants causes a constitutive transcription of PDF1.2. When grown in soil, the transgenic plants exhibited an increased root development compared with WT plants and an enhanced resistance toward the root pathogen Pythium irregulare. Six paralogs of PROPEP1 are present in Arabidopsis, and orthologs have been identified in species of several agriculturally important plant families, where they are of interest for their possible use in crop improvement.

Phenazine Compounds in Fluorescent Pseudomonas Spp. Biosynthesis and Regulation

Abstract: The phenazines include upward of 50 pigmented, heterocyclic nitrogen-containing secondary metabolites synthesized by some strains of fluorescent Pseudomonas spp. and a few other bacterial genera. The antibiotic properties of these compounds have been known for over 150 years, but advances within the past two decades have provided significant new insights into the genetics, biochemistry, and regulation of phenazine synthesis, as well as the mode of action and functional roles of these compounds in the environment. This new knowledge reveals conservation of biosynthetic enzymes across genera but raises questions about conserved biosynthetic mechanisms, and sets the stage for improving the performance of phenazine producers used as biological control agents for soilborne plant pathogens.

Costs of a predictable switch between simple cognitive tasks following severe closed-head injury

Abstract: The authors used a predictable, externally cued task-switching paradigm to investigate executive control in a severe closed-head injury (CHI) population. Eighteen individuals with severe CHI and 18 controls switched between classifying whether a digit was odd or even and whether a letter was a consonant or vowel on every 4th trial. The target stimuli appeared in a circle divided into 8 equivalent parts. Presentation of the stimuli rotated clockwise. Participants performed the switching task at both a short (200 ms) and a long (1,000 ms) preparatory interval. Although the participants with CHI exhibited slower response times and greater switch costs, similar to controls, additional preparatory time reduced the switch costs, and the switch costs were limited to the 1st trial in the run. These findings indicate that participants with severe CHI were able to take advantage of time to prepare for the task switch, and the executive control processes involved in the switch costs were completed before the 1st trial of the run ended.

Endocrine Disruptor Vinclozolin Induced Epigenetic Transgenerational Adult-Onset Disease

Abstract: The fetal basis of adult disease is poorly understood on a molecular level and cannot be solely attributed to genetic mutations or a single etiology. Embryonic exposure to environmental compounds has been shown to promote various disease states or lesions in the first generation (F1). The current study used the endocrine disruptor vinclozolin (antiandrogenic compound) in a transient embryonic exposure at the time of gonadal sex determination in rats. Adult animals from the F1 generation and all subsequent generations examined (F1–F4) developed a number of disease states or tissue abnormalities including prostate disease, kidney disease, immune system abnormalities, testis abnormalities, and tumor development (e.g. breast). In addition, a number of blood abnormalities developed including hypercholesterolemia. The incidence or prevalence of these transgenerational disease states was high and consistent across all generations (F1–F4) and, based on data from a previous study, appears to be due in part to epig...

Epigenetic Transgenerational Actions of Endocrine Disruptors

Abstract: Endocrine disruptors have recently been shown to promote an epigenetic transgenerational phenotype involving a number of disease states (e.g. male infertility). The anti-androgenic fungicide vinclozolin was found to act transiently at the time of embryonic sex determination to promote in the F1 generation a spermatogenic cell defect and subfertility in the male. When the animals were allowed to age up to 1 yr, a number of other disease states developed. This phenotype was transferred through the male germ line to all subsequent generations analyzed (F1-F4). The ability of an environmental factor (i.e. endocrine disruptor) to promote an epigenetic transgenerational phenotype impacts the potential hazards of environmental toxins, mechanisms of disease etiology, and evolutionary biology. The biological importance of the epigenetic actions of environmental agents is reviewed in the context of the primordial germ cell and development of epigenetic transgenerational phenotypes.

Spiropyran-based photochromic polymer nanoparticles with optically switchable luminescence.

Abstract: Polymer nanoparticles of 40−400 nm diameter with spiropyran−merocyanine dyes incorporated into their hydrophobic cavities have been prepared; in contrast to their virtually nonfluorescent character in most environments, the merocyanine forms of the encapsulated dyes are highly fluorescent. Spiro−mero photoisomerization is reversible, allowing the fluorescence to be switched “on” and “off” by alternating UV and visible light. Immobilizing the dye inside hydrophobic pockets of nanoparticles also improves its photostability, rendering it more resistant than the same dyes in solution to fatigue effects arising from photochemical switching. The photophysical characteristics of the encapsulated fluorophores differ dramatically from those of the same species in solution, making nanoparticle-protected hydrophobic fluorophores attractive materials for potential applications such as optical data storage and switching and biological fluorescent labeling. To evaluate the potential for biological tagging, these optica...

Colloid transport of plutonium in the far-field of the Mayak Production Association, Russia.

Abstract: Sorption of actinides, particularly plutonium, onto submicrometer-sized colloids increases their mobility, but these plutonium colloids are difficult to detect in the far-field. We identified actinides on colloids in the groundwater from the Mayak Production Association, Urals, Russia; at the source, the plutonium activity is ∼1000 becquerels per liter. Plutonium activities are still 0.16 becquerels per liter at a distance of 3 kilometers, where 70 to 90 mole percent of the plutonium is sorbed onto colloids, confirming that colloids are responsible for the long-distance transport of plutonium. Nano–secondary ion mass spectrometry elemental maps reveal that amorphous iron oxide colloids adsorb Pu(IV) hydroxides or carbonates along with uranium carbonates.

The ACS Virgo Cluster Survey. VI. Isophotal Analysis and the Structure of Early-Type Galaxies

Abstract: (Abridged) We present a detailed analysis of the morphology, isophotal parameters and surface brightness profiles for 100 early-type members of the Virgo Cluster, from dwarfs (M_B = -15.1 mag) to giants (M_B = -21.8 mag). Each galaxy has been imaged in two filters, closely resembling the Sloan g and z passbands, using the Advanced Camera for Surveys on board the Hubble Space Telescope. Dust and complex morphological structures are common, with kiloparsec-scale stellar disks, bars, and nuclear stellar disks seen in 60% of galaxies with intermediate luminosity (-20 < M_B < -17), and dust seen in 42% of galaxies brighter than M_B = -18.9 mag. Dust morphologies range from faint wisps and patches on tens of parsec scales, to regular, highly organized kpc-scale dust disks, often showing evidence of recent star formation. Surface brightness profiles and isophotal parameters are derived typically within 8 kpc from the center for the brightest galaxies, and 1.5 kpc for the faintest systems, with a resolution (FWHM) of 7 pc. Based on a parametrization of the surface brightness profiles in terms of a Sersic or core-Sersic model, we find that 1) there is no evidence of a bimodal behavior of the slope, gamma, of the profile in the innermost regions; 2) although the brightest galaxies have shallow inner profiles, the shallowest profiles (lowest gamma values) are found in faint dwarf systems; 3) the widely adopted separation of early-type galaxies between "core" and "power-law" types, which had originally been prompted by the claim of a clear bimodal distribution of gamma values, is untenable; and 4) there is no evidence of a structural dichothomy between dwarf and regular ellipticals.

CHAPTER 38 – Puberty in the Rat

Abstract: This chapter discusses the modulatory influences regulating the timing of puberty in rats. The developmental process that leads to puberty in the female rat is based on an extraordinarily complex series of interrelated events. The central nervous system (CNS) plays a critical role by controlling both anterior pituitary function, through the secretion of hypothalamic factors, and the ovary via pituitary hormones and direct neural inputs. The central neuroendocrine regulation of luteinizing hormone-releasing hormone (LHRH) production and gonadotropin secretion initiates the onset of puberty in rats through LH actions on the Leydig cells to increase androgen production and follicle-stimulating hormone (FSH) actions on the Sertoli cell to enhance Leydig cell function and initiate Sertoli cell differentiation needed for the induction of spermatogenesis. The negative feedback of androgens and inhibin regulate gonadotropin production during the progression of puberty to the adult stage. The coordinated control of this hypothalamic–pituitary axis with the testis involves molecular events at the LHRH neuron and pituitary levels and at the testis somatic cell level. The resulting endocrine events control the onset and progression of puberty in the rat.

A Fundamental Relation between Compact Stellar Nuclei, Supermassive Black Holes, and Their Host Galaxies

Abstract: Imaging surveys with the Hubble Space Telescope (HST) have shown that ?50%-80% of low- and intermediate-luminosity galaxies contain a compact stellar nucleus at their center, regardless of host galaxy morphological type. We combine HST imaging for early-type galaxies from the ACS Virgo Cluster Survey with ground-based long-slit spectra from KPNO to show that the masses of compact stellar nuclei in Virgo Cluster galaxies obey a tight correlation with the masses of the host galaxies. The same correlation is obeyed by the supermassive black holes (SBHs) found in predominantly massive galaxies. The compact stellar nuclei in the Local Group galaxies M33 and NGC 205 are also found to fall along this same scaling relation. These results indicate that a generic by-product of galaxy formation is the creation of a central massive object (CMO)?either an SBH or a compact stellar nucleus?that contains a mean fraction, ?0.2%, of the total galactic mass. In galaxies with masses greater than gal ~ a few × 1010 ?, SBHs appear to be the dominant mode of CMO formation.

Biofluidmechanics of Reproduction

Abstract: Mammalian fertilization requires the coordinated activity of motile spermatozoa, muscular contractions of the uterus and oviduct, as well as ciliary beating. These elastic structures generate forces that drive fluid motion, but their configurations are, in turn, determined by the fluid dynamics. We review the basic fluid mechanical aspects of reproduction, including flagellar/ciliary beating and peristalsis. We report on recent biological studies that have shed light on the relative importance of the mechanical ingredients of reproduction. In particular, we examine sperm motility in the reproductive tract, ovum pickup and transport in the oviduct, as well as sperm-egg interactions. We review recent advances in understanding the internal mechanics of cilia and flagella, flagellar surface interaction, sperm motility in complex fluids, and the role of fluid dynamics in embryo transfer. We outline promising computational fluid dynamics frameworks that may be used to investigate these complex, fluid-structure interactions.

A longitudinal study of relational aggression, physical aggression, and children's social-psychological adjustment.

Abstract: Although great strides have recently been made in our understanding of relational aggression and its consequences, one significant limitation has been the lack of prospective studies. The present research addressed this issue by identifying and assessing groups of relationally aggressive, physically aggressive, relationally plus physically aggressive (co-morbid), and nonaggressive children during their third grade year in elementary school and then reassessing them a year later, during fourth-grade (N = 224, 113 girls). Two aspects of social-psychological adjustment were assessed during both assessment periods including internalizing difficulties (i.e., withdrawal, depression/anxiety, and somatic complaints) and externalizing problems (i.e., aggressive behavior, delinquency). It was revealed that the strongest predictor of future social-psychological adjustment problems and increases in these problems from third to fourth was the combination of relational and physical aggression. Relational aggression also contributed unique information, relative to physical aggression, in the prediction of future maladjustment. Implications of these findings for future research and prevention efforts, particularly for aggressive girls, are discussed.

Taxol Biosynthesis and Molecular Genetics

Abstract: Biosynthesis of the anticancer drug Taxol in Taxus (yew) species involves 19 steps from the universal diterpenoid progenitor geranylgeranyl diphosphate derived by the plastidial methyl erythritol phosphate pathway for isoprenoid precursor supply. Following the committed cyclization to the taxane skeleton, eight cytochrome P450-mediated oxygenations, three CoA-dependent acyl/aroyl transfers, an oxidation at C9, and oxetane (D-ring) formation yield the intermediate baccatin III, to which the functionally important C13-side chain is appended in five additional steps. To gain further insight about Taxol biosynthesis relevant to the improved production of this drug, and to draw inferences about the organization, regulation, and origins of this complex natural product pathway, Taxus suspension cells (induced for taxoid biosynthesis by methyl jasmonate) were used for feeding studies, as the foundation for cell-free enzymology and as the source of transcripts for cDNA library construction and a variety of cloning strategies. This approach has led to the elucidation of early and late pathway segments, the isolation and characterization of over half of the pathway enzymes and their corresponding genes, and the identification of candidate cDNAs for the remaining pathway steps, and it has provided many promising targets for genetically engineering more efficient biosynthetic production of Taxol and its precursors.