Detailed Fate Research Articles

Effect of different oligomerization assemblies of γ-conglutin on its interaction behavior with vitexin.

Several different factors underlie the molecular mechanisms of phenolic compound-protein interactions. They include the environmental conditions. In the case of γ-conglutin, pH conditions translate directly into the adoption of two distinct oligomeric assemblies, i.e. hexameric (pH 7.5) or monomeric (pH 4.5). This paper reports research on the pH-dependent oligomerization of γ-conglutin in terms of its ability to form complexes with a model flavonoid (vitexin). Fluorescence-quenching thermodynamic measurements indicate that hydrogen bonds, electrostatic forces, and van der Waals interactions are the main driving forces involved in the complex formation. The interaction turned out to be a spontaneous and exothermic process. Assessment of structural composition (secondary structure changes and arrangement/dynamics of aromatic amino acids), molecular size, and the thermal stability of the different oligomeric forms showed that γ-conglutin in a monomeric state was less affected by vitexin during the interaction. The data show precisely how environmental conditions might influence phenolic compound-protein complex formation directly. This knowledge is essential for the preparation of food products containing γ-conglutin. The results can contribute to a better understanding of the detailed fate of this unique health-promoting lupin seed protein after its intake. © 2023 Society of Chemical Industry.

Pharmacokinetics and residue elimination of metalaxyl enantiomers in snakehead (Channa argus)

Metalaxyl is widely used as a racemic mixture to control various plant and aquatic animal diseases caused by fungal infection. It has been proved that the two enantiomers of metalaxyl have significant differences in activity and toxicity, while little is known about the pharmacokinetics and residual elimination of the individual enantiomers of metalaxyl in aquatic animals. In this study, a simple, sensitive and reliable chiral liquid chromatography-tandem mass spectrometry method was established for the determination of metalaxyl and its metabolite metalaxyl acid enantiomers, which was applied to study the stereoselective pharmacokinetics and residue elimination of individual enantiomers in snakehead (Channa argus). The snakehead was bath administered with the racemic metalaxyl and the highly active enantiomeric R-metalaxyl, respectively. The results showed that the elimination of metalaxyl in the plasma and tissues of snakehead was enantioselective. The concentration of metalaxyl in tissues was kidney > liver > muscle, and preferential degradation of the R-metalaxyl enantiomer was observed in three tissues. However, no transformation between metalaxyl enantiomers was observed in snakehead plasma and tissues when R-metalaxyl enantiomer was administered. In addition, the distribution of metalaxyl acid enantiomers in tissues is closely related to the parent compound metalaxyl, and S-metalaxyl acid was only observed in the kidney and slowly eliminated. The highest concentrations of both metalaxyl and metalaxyl acid were observed in the kidney, suggesting that kidney is the major site of metalaxyl metabolism in snakehead. Given that the highly active R-metalaxyl has a shorter half-life and does not undergo transformation between enantiomers in snakehead, it may be a better choice to use R-metalaxyl instead of racemic metalaxyl in clinical practice. The results are helpful to understand the detailed fate of metalaxyl enantiomers and should be considered in future aquaculture, environmental pollution risk and food safety evaluations.

Hydrodynamic Impacts on the Fate of Polychlorinated Biphenyl 153 in the Marine Environment

Due to their long half-life, polychlorinated biphenyls (PCBs) tend to contaminate not only coastal areas, but they travel over long distances, eventually reaching remote areas such the Arctic. The physical and biogeochemical features of every coastal area govern the main distribution patterns of freshly introduced PCBs into the marine system. Some of these processes are determined by chemical properties of the individual PCB congener. Thus, atmospheric influx along with ad/absorption on non-living organic material, photolytical and biological degradation processes vary from one PCB congener to another. For a detailed fate analysis of individual congeners, we developed a new chemical model, based on the GOTM-ECOSMO-FABM model framework. Here, we exemplarily present results for PCB153 based on 1D simulations of four regions in the North-Baltic Sea. The study area is characterized by different hydrodynamic and biogeochemical conditions. We investigate the impact of resuspension, mixing and the biological pump, sea ice and tides on the final phasal distribution of PCB153. Different combinations of these factors lead to the development of different areas of PCB153 accumulation, with the formation of hotspot areas, and influence the total uptake and concentration of PCB153 in the water column. As a result, two major dynamics determine the fate of PCB153 in the coastal ocean: (i) Primary production leads to PCB153 being adsorbed by organic material. Partitioning to organic material and sedimentation of organic particles removes dissolved PCB153 from the surface ocean and increases atmospheric influx. (ii) Tidal-induced resuspension and mixing control the benthic–pelagic exchange of PCB153 and its distribution in the water column. Depending on the resuspension regime and stratification, sediments can become a permanent (Gotland Deep, the Baltic Sea) or seasonal sink for PCB153. In regions with seasonal stratification and high near bottom turbulence (Northern North Sea), resuspension events can lead to pronounced peaks in PCB153 concentrations and can therefore have a major impact on bioaccumulation. Under the conditions of permanent mixing and high bottom turbulence (Southern North Sea, Bothnian Bay), pollutants are hardly accumulating in sediments.

Microcosm test for pesticide fate assessment in planted water filters: 13C,15N-labeled glyphosate as an example

Planted filters are often used to remove pesticides from runoff water. However, the detailed fate of pesticides in the planted filters still remains elusive. This hampers an accurate assessment of environmental risks of the pesticides related to their fate and thereby development of proper mitigation strategies. In addition, a test system for the chemical fate analysis including plants and in particular for planted filters is not well established yet. Therefore, we developed a microcosm test to simulate the fate of pesticide in planted filters, and applied 2-13C,15N-glyphosate as a model pesticide. The fate of 2-13C,15N-glyphosate in the planted microcosms over 31 day-incubation period was balanced and compared with that in the unplanted microcosms. The mass balance of 2-13C,15N-glyphosate turnover included 13C mineralization, degradation products, and the 13C and 15N incorporation into the rhizosphere microbial biomass and plants. We observed high removal of glyphosate (> 88%) from the water mainly due to adsorption on gravel in both microcosms. More glyphosate was degraded in the planted microcosms with 4.1% of 13C being mineralized, 1.5% of 13C and 3.8% of 15N being incorporated into microbial biomass. In the unplanted microcosms, 1.1% of 13C from 2-13C,15N-glyphosate was mineralized, and only 0.2% of 13C and 0.1% of 15N were assimilated into microbial biomass. The total recovery of 13C and 15N was 81% and 85% in planted microcosms, and 91% and 93% in unplanted counterparts, respectively. The microcosm test was thus proven to be feasible for mass balance assessments of the fate of non-volatile chemicals in planted filters. The results of such studies could help better manage and design planted filters for pesticide removal.

Biological treatment of Hydrothermal Liquefaction (HTL) wastewater: Analytical evaluation of continuous process streams

In order to deal with hydrothermal liquefaction wastewater (HTWW), a new anaerobic-aerobic continuous process was developed. The process, which included a sequence of Up-Flow Anaerobic Sludge Blanket (UASB) and downstream aerobic Continuously Stirred Tank Reactor (CSTR), was tested on the HTWW obtained from Waste to Fuel® demo plant developed by ENI s.p.a. [1]. Performance of the system was evaluated in term of methane yield and chemical oxygen demand (COD) abatement capability. Detailed fate of organic compounds was evaluated through different analytical techniques, highlighting main issues and potential of HTWW biological treatment. The system was fed with neat HTWW (189 gCOD L−1) for 2.5 y, with variable organic loading rate (OLR) and minimal external inputs. UASB reactors converted most of HTWW organics into volatile fatty acids (VFA) and methane with concurrent precipitation of oily like insoluble, whereas aerobic CSTR removed VFA from anaerobic effluent. Under regime conditions (ORL equal to 0.5 gCOD L−1 d−1) COD decreased from 189 to 6.6 gCOD L-1, showing 97 % COD abatement with the coupled anaerobic-aerobic treatment. Such a COD abatement was obtained by means of multiple effects, namely biomethanation, precipitation of organic matter and aerobic oxidation of fermentation products (VFA produced in anaerobic digestion) and subsequent aerobic oxidation (in downstream aerobic reactor). These effects accounted for 43, 40 and 17 % of the total COD decrease, respectively. Inhibition phenomena were the key challenge for improving methane yields and system productivities. The overall results confirmed that valorization of HTWW is a feasible task, albeit rather challenging.

Treatment characteristics of various sediment components spiked with 2-chlorobiphenyl using reactive activated carbon

Previously, the concept of reactive activated carbon (RAC), where the porous structure of activated carbon (AC) is impregnated with palladized zerovalent iron, has been proposed to be effective to adsorb and dechlorinate polychlorinated biphenyls (PCBs). To explain the low dechlorination of PCBs bound to actual aquatic sediments under remediation with RAC, this study investigated the role of various solid organic and inorganic sediment components in adsorbing and desorbing PCBs. Detailed fate and transport mechanism of 2-chlorinated biphenyl (2-ClBP) spiked to sediment components, including kaolin, montmorillonite (MMT), coal, graphite, AC, and their mixture, was revealed. Adsorption and holding capability of sediment components toward 2-ClBP strongly influenced amount of spiked 2-ClBP, amount of desorbed 2-ClBP, overall dechlorination of 2-ClBP to biphenyl (BP), and eventual partitioning of 2-ClBP and BP to water, sediment component, and RAC. Order of the amount of spiked 2-ClBP to sediment components after drying, following AC > mixture > coal > graphite > kaolin > MMT, was in agreements (in opposite direction) with order of the amount of desorbed 2-ClBP and order of overall 2-ClBP dechlorination. Substantial role of organic components in aquatic sediments for holding 2-ClBP and thus preventing it from dechlorination on RAC was proven.

Dense shelf water spreading from Antarctic coastal polynyas to the deep Southern Ocean: A regional circumpolar model study

AbstractThe spreading of dense shelf water (DSW) from Antarctic coastal margins to lower latitudes plays a vital role in the ocean thermohaline circulation and the global climate system. Through enhanced localized sea ice production in Antarctic coastal polynyas, cold and saline DSW is formed over the continental shelf regions as a precursor to Antarctic Bottom Water (AABW). However, the detailed fate of coastal DSW over the Southern Ocean is still unclear. Here we conduct extensive passive tracer experiments using a circumpolar ocean‐sea ice‐ice shelf model to investigate pathways of the regional polynya‐based DSW from the Antarctic margins to the deep Southern Ocean basins. In the numerical experiments, the Antarctic coastal margin is divided into nine regions, and a passive tracer is released from each region at the same rate as the local sea ice production. The modeled spatial distribution of the total concentration of the nine tracers is consistent with the observed AABW distribution and clearly demonstrates nine routes of the DSW over the Southern Ocean along its bottom topography. Furthermore, the model shows that while ∼50% of the total tracer is distributed northward from the continental shelf to the deep ocean, ∼7% is transported poleward beneath ice shelf cavities. The comprehensive tracer experiments allow us to estimate the contribution of local DSW to the total concentration along each of the pathways.

Glyphosate and AMPA in the estuaries of the Baltic Sea method optimization and field study

Water samples from ten German Baltic estuaries were collected in 2012 in order to study the presence of the herbicide glyphosate, its primary metabolite AMPA and their potential transport to the marine environment. For the analyses an LC–MS/MS based analytical method after derivatization with FMOC-Cl was optimized and validated for marine water samples. All investigated estuarine stations were contaminated with AMPA and nine of them also with glyphosate. Concentration ranges observed were 28 to 1690ng/L and 45 to 4156ng/L for glyphosate and AMPA, respectively with strong spatial and temporal fluctuations. Both contaminants were found at inbound sampling sites in the stream Muehlenfliess and concentrations decreased along the salinity gradient to the estuaries of the Baltic Sea. The data obtained in this study clearly depict the transport of glyphosate and AMPA to the Baltic Sea. Hence, detailed fate and risk assessment for both contaminants in marine environments are required.

Evolution of carbon isotope signatures during reactive transport of hydrocarbons in heterogeneous aquifers

Compound-specific isotope analysis (CSIA) of organic pollutants has become a well-established tool for assessing the occurrence and extent of biodegradation processes in contaminated aquifers. However, the precision of CSIA is influenced by the degree to which assumptions underlying CSIA data interpretation hold under realistic field-scale conditions. For the first time this study demonstrates how aquifer analogs combined with reactive transport models offer an underexplored way to develop generic process understanding, evaluate monitoring and quantification strategies in highly heterogeneous subsurface settings. Data from high-resolution aquifer analogs were used in numerical experiments to track the propagation of a representative oxidizable organic compound (toluene) within a variety of realistic heterogeneous aquifers and to investigate its detailed fate. The simulations were used to analyze (1) the effects of physical aquifer heterogeneities on spatiotemporal patterns of contaminant concentrations and isotope signatures, (2) the performance of the commonly applied Rayleigh equation and (3) the applicability of an extension of the Rayleigh equation for complex hydrogeological conditions. The results indicate that if field-derived enrichment factors are applied without corrections for dilution, the conventional Rayleigh equation is inaccurate and estimates for biodegradation are typically overestimated and unreliable in heterogeneous aquifers. Underestimations can occur due to the partial source zone depletion. In contrast, if dilution can be accurately accounted for, field-derived enrichment factors comprise a suitable alternative to laboratory-derived and redox-specific enrichment factors. The study also examines to what extent variations in monitoring/sampling strategies influence the obtained results. Especially measurements from long-screened wells (>1m) reveal to be inappropriate for the application of the Rayleigh equation in the investigated aquifer analogs, as low resolution data sampled from the simulated scenarios only enable a qualitative assessment of biodegradation. Measurements from both long- and short-screened wells employing the Rayleigh equation streamline approach are only partly viable for in situ biodegradation measurements in heterogeneous systems.

Fate mapping of gallbladder progenitors in posteroventral foregut endoderm of mouse early somite-stage embryos.

In early embryogenesis, the posteroventral foregut endoderm gives rise to the budding endodermal organs including the liver, ventral pancreas and gallbladder during early somitogenesis. Despite the detailed fate maps of the liver and pancreatic progenitors in the mouse foregut endoderm, the exact location of the gallbladder progenitors remains unclear. In this study, we performed a DiI fate-mapping analysis using whole-embryo cultures of mouse early somite-stage embryos. Here, we show that the majority of gallbladder progenitors in 9–11-somite-stage embryos are located in the lateral-most domain of the foregut endoderm at the first intersomite junction level along the anteroposterior axis. This definition of their location highlights a novel entry point to understanding of the molecular mechanisms of initial specification of the gallbladder.

Endocytic Traffic: Vesicle Fusion Cascade in the Early Endosomes

New research shows that vesicles in the early endosomal network coalesce according to a classical theoretical description of aggregation put forward by Smoluchowski more than 100 years ago. This gives a new tool for unraveling complexities of the endocytic pathways.

Aquatic Risk Assessment of Enzymes used in Household Detergents

Abstract Exposure and effect assessment including risk characterisation of enzymes used in household detergents were conducted. Predicted environmental concentrations were calculated by use of traditional methods and detailed transport and fate modelling. Predicted no effect concentrations were derived from the results of ecotoxicological studies conducted with subtilisin and other available information on the acute and chronic aquatic toxicity. Risk characterisation ratios were defined as the ratio between the predicted environmental concentration and the predicted no effect concentration. Risk characterisation ratios were calculated for fresh and marine water and were found to be between 0.0002 and 0.06, i.e. all well below 1. This shows that under the given conditions, the use of enzymes in household detergents does not present a risk to the aquatic environment.

Aquatic Risk Assessment of Alcohol Ethoxylates, Alcohol Ethoxysulphates and Linear Alkylbenzene Sulphonate used in Household Detergents

Abstract Exposure and effect assessments including risk characterisation of the surfactants alcohol ethoxylates, alcohol ethoxysulphates and linear alkylbenzene sulphonates used in household detergents were conducted. Predicted environmental concentrations were calculated by use of traditional methods and detailed transport and fate modelling. Predicted no effect concentrations were derived from the available information on the acute and chronic aquatic toxicity. Risk characterisation ratios were defined as the ratio of the predicted environmental concentration to the predicted no effect concentration. Risk characterisation ratios were calculated for fresh water and marine water and were below 1. This indicates that based on the methods employed in this study the use of alcohol ethoxylates, alcohol ethoxysulphates and linear alkylbenzene sulphonate in household detergents does not appear to present a risk to the aquatic environment.

The fate of dividing cells during lens morphogenesis, differentiation and growth

Early in development, the ocular lens establishes its distinctive architecture, and this is maintained throughout life as the lens continues to grow. This growth is tightly regulated through the proliferation of the lens epithelial cells and their subsequent differentiation into specialized elongated fiber cells. Although much work has been carried out to define these patterns of growth, very little has been reported on the detailed fate and kinetics of lens cells during embryogenesis. Using BrdU-incorporation, the present study has attempted to follow the fate of lens cells that have undergone at least one round of DNA synthesis during the early stages of lens morphogenesis. Results from this work have confirmed that the rate of lens cell proliferation and new fiber cell differentiation progressively slows as the lens differentiates and grows. In addition, these studies have shown that early in lens development, not all DNA synthesis is restricted to the lens epithelium, with some elongating fiber cells retaining the ability to undergo DNA synthesis. Adopting this system we have also been able to place the initiation of secondary fiber cell differentiation in the mouse lens by E12.5, concomitant with the loss of the lens vesicle lumen by the elongating primary fiber cells. Overall, this study has allowed us to revisit some of the mechanisms involved in early lens development, has provided us with insights into the fate of cells during this rapid phase of murine lens growth, and has provided a novel method to study the rate of new fiber cell differentiation over a defined period of lens development and growth.

Phylogenetic analysis of developmental and postnatal mouse cell lineages

Fate maps depict how cells relate together through past lineage relationships, and are useful tools for studying developmental and somatic processes. However, with existing technologies, it has not been possible to generate detailed fate maps of complex organisms such as the mouse. We and others have therefore proposed a novel approach, "phylogenetic fate mapping," where patterns of somatic mutation carried by the individual cells of an animal are used to retrospectively deduce lineage relationships through phylogenetic inference. Here, we have cataloged genomic polymorphisms at 324 mutation-prone polyguanine tracts for nearly 300 cells isolated from a single mouse, and have explored the cells' lineage relationships both phylogenetically and through a network-based approach. We present a model of mouse embryogenesis, where an early period of substantial cell mixing is followed by more coherent growth of clones later. We find that cells from certain tissues have greater numbers of close relatives in other specific tissues than expected from chance, suggesting that those populations arise from a similar pool of ancestral lineages. Finally, we have investigated the dynamics of cell turnover (the frequency of cell loss and replacement) in postnatal tissues. This work offers a longitudinal study of developmental lineages, from conception to adulthood, and provides insight into basic questions of mouse embryology as well as the somatic processes that occur after birth.

下水道未整備地域を流れる小河川における生活関連汚染物質の動態解明と濃度簡易予測モデルの構築〜徳島市冷田川を例として

An antifungal agent triclosan (TCS) and an anionic surfactant linear alkylbenzene sulfonate (LAS) were listed as candidates of detailed ecotoxicological risk assessment by Japan Ministry of Environment. These two compounds were selected to investigate the detailed fate and transport of these household effluent-originated pollutants in a small urban stream, Tsumeta Creek, Tokushima, Japan with no sewer system. Laboratory biodegradation and photodegradation experiments were conducted to determine first-order rate constants. Using these parameters and the environmental loading estimated based on PRTR or other statistics, a simple numerical model was constructed to estimate aquatic concentrations of TCS and C12-LAS in each sampling point in Tsumeta Creek and these values were compared with the measured values. Overall, the estimated values were acceptable and were in the range of 50 to 200% of the measured ones. The results suggest the concentration of TCS and C12-LAS is in the level of potential effects on aquatic organisms. From the viewpoint of the conservation of ecosystem, the mechanism of aquatic pollution and the removal of environmental loading of household effluent-originated pollutants such as TCS and LAS need to be further investigated in the unsewered area using both model prediction and monitoring.

Origin of the zebrafish endocrine and exocrine pancreas

Here, we report a detailed fate map of the zebrafish pancreas at the early gastrula stage of development (6 hours postfertilization; hpf). We show that, at this stage, both pancreas and liver progenitors are symmetrically localized in two broad domains relative to the dorsal organizer. We demonstrate that the dorsal and ventral pancreatic buds can derive from common progenitor pools at 6 hpf, but often derive from independent populations. Endocrine vs. exocrine pancreas show a similar pattern of progenitors, consistent with descriptions of the dorsal bud being strictly endocrine and the ventral bud primarily exocrine. In general, we find that endocrine/dorsal bud progenitors are located more dorsally than the exocrine pancreas/ventral bud progenitors. Later in gastrulation (10 hpf), pancreas progenitors have migrated to bilateral domains at the equator of the embryo. Our fate map will assist with design and interpretation of future experiments to understand early pancreas development.

Specification of cell fate along the proximal-distal axis in the developing chick limb bud

Pattern formation along the proximal-distal (PD) axis in the developing limb bud serves as a good model for learning how cell fate and regionalization of domains, which are essential processes in morphogenesis during development, are specified by positional information. In the present study, detailed fate maps for the limb bud of the chick embryo were constructed in order to gain insights into how cell fate for future structures along the PD axis is specified and subdivided. Our fate map revealed that there is a large overlap between the prospective autopod and zeugopod in the distal limb bud at an early stage (stage 19), whereas a limb bud at this stage has already regionalized the proximal compartments for the prospective stylopod and zeugopod. A clearer boundary of cell fate specifying the prospective autopod and zeugopod could be seen at stage 23, but cell mixing was still detectable inside the prospective autopod region at this stage. Detailed analysis of HOXA11 AND HOXA13 expression at single cell resolution suggested that the cell mixing is not due to separation of some different cell populations existing in a mosaic. Our findings suggest that a mixable unregionalized cell population is maintained in the distal area of the limb bud, while the proximal region starts to be regionalized at the early stage of limb development.

Pharmaceuticals, Personal Care Products, and Endocrine Disruptors in Water: Implications for the Water Industry

For over 70 years, scientists have reported that certain synthetic and natural compounds could mimic natural hormones in the endocrine systems of animals. These substances are now collectively known as endocrine-disrupting compounds (EDCs), and have been linked to a variety of adverse effects in both humans and wildlife. More recently, pharmaceuticals and personal care products (PPCPs) have been discovered in various surface and ground waters, some of which have been linked to ecological impacts at trace concentrations. The majority of EDCs and PPCPs are more polar than traditional contaminants and several have acidic or basic functional groups. These properties, coupled with occurrence at trace levels (i.e., <1 μg/L), create unique challenges for both removal processes and analytical detection. Reports of EDCs and PPCPs in water have raised substantial concern among the public and regulatory agencies; however, very little is known about the fate of these compounds during drinking and wastewater treatment. Numerous studies have shown that conventional drinking and wastewater treatment plants can not completely remove many EDCs and PPCPs. Oxidation with chlorine and ozone can result in transformation of some compounds with reactive functional groups under the conditions employed in water and wastewater treatment plants. Advanced treatment technologies, such as activated carbon and reverse osmosis, appear viable for the removal of many trace contaminants including EDCs and PPCPs. Future research needs include more detailed fate and transport data, standardized analytical methodology, predictive models, removal kinetics, and determination of the toxicological relevance of trace levels of EDCs and PPCPs in water.

Epiblast and primitive-streak origins of the endoderm in the gastrulating chick embryo.

Gastrulation is characterized by the extensive movements of cells. Fate mapping is used to follow such cell movements as they occur over time, and prospective fate maps have been constructed for several stages of the model organisms used in modern studies in developmental biology. In chick embryos, detailed fate maps have been constructed for both prospective mesodermal and ectodermal cells. However, the origin and displacement of the prospective endodermal cells during crucial periods in gastrulation remain unclear. This study had three aims. First, we determined the primitive-streak origin of the endoderm using supravital fluorescent markers, and followed the movement of the prospective endodermal cells as they dispersed to generate the definitive endodermal layer. We show that between stages 3a/b and 4, the intraembryonic definitive endoderm receives contributions mainly from the rostral half of the primitive streak, and that endodermal movements parallel those of ingressing adjacent mesodermal subdivisions. Second, the question of the epiblast origin of the endodermal layer was addressed by precisely labeling epiblast cells in a region known to give rise to prospective somitic cells, and following their movement as they underwent ingression through the primitive streak. We show that the epiblast clearly contributes prospective endodermal cells to the primitive streak, and subsequently to definitive endoderm of the area pellucida. Finally, the relationship between the hypoblast and the definitive endoderm was defined by following labeled rostral primitive-streak cells over a short period of time as they contributed to the definitive endoderm, and combining this with in situ hybridization with a riboprobe for Crescent, a marker of the hypoblast. We show that as the definitive endodermal layer is laid down, there is cell-cell intercalation at its interface with the displaced hypoblast cells. These data were used to construct detailed prospective fate maps of the endoderm in the chick embryo, delineating the origins and migrations of endodermal cells in various rostrocaudal levels of the primitive streak during key periods in early development.