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Poultry Science May 2014Egg producers in the United States are utilizing a variety of commercial egg production systems to provide consumer choice and meet legislative requirements. Consumer...
Egg producers in the United States are utilizing a variety of commercial egg production systems to provide consumer choice and meet legislative requirements. Consumer egg grades in the United States were developed for conventional cage production, and it is unclear what effect alternative production systems might have on egg quality during retail and consumer home storage. The current study was undertaken to determine what changes in egg quality characteristics occur during extended cold storage for commercially produced conventional cage, enriched colony cage, and cage-free aviary eggs. During 12 wk of cold storage, egg weight, albumen height, Haugh unit, static compression shell strength, vitelline membrane strength and deformation, yolk index, shell dynamic stiffness, and whole egg total solids were monitored. Overall, aviary and enriched eggs were significantly (P < 0.05) heavier than conventional cage. Albumen height and Haugh unit (P < 0.05) were significantly greater for conventional cage than enriched eggs. Static compression shell strength was greatest (P < 0.05) for enriched eggs compared with aviary. No overall housing system effects for yolk measurements, shell dynamic stiffness, or whole egg total solids were observed. Albumen height, Haugh unit, and yolk quality measurements were all greatest at 0 and lowest at 12 wk of storage (P < 0.05). The rate of quality change among the housing systems for each measured attribute at 4, 6, and 12 wk was determined. Other than differences in the change of egg weight at 4 wk, no significant differences in the rate of quality decline were found among the housing systems. The results of the current study indicate that current US egg quality standards should effectively define quality for commercially produced conventional cage, enriched colony cage, and cage-free aviary eggs.
Topics: Animal Husbandry; Animals; Chickens; Cold Temperature; Female; Food Storage; Housing, Animal; Ovum
PubMed: 24795324
DOI: 10.3382/ps.2013-03631 -
Current Protocols in Toxicology Feb 2014The fruit fly (Drosophila melanogaster) has long been a premier model for developmental biologists and geneticists. In toxicology studies, Drosophila has only recently...
The fruit fly (Drosophila melanogaster) has long been a premier model for developmental biologists and geneticists. In toxicology studies, Drosophila has only recently gained broader recognition as a tool to elaborate molecular genetic mechanisms of toxic substances. In this article, two practical applications of Drosophila for developmental toxicity assays are described. The first assay takes advantage of newly developed methods to render the fly embryo accessible to small molecules, toxicants, and drugs. The second assay engages straightforward exposures to developing larvae and easy-to-score outcomes of adult development. With the extensive collections of flies that are publicly available and the ease of creating transgenic flies, these two assays have a unique power for identifying and characterizing molecular mechanisms and cellular pathways specific to the mode of action of a number of toxicants and drugs.
Topics: Animals; Drosophila; Larva; Models, Biological
PubMed: 24789363
DOI: 10.1002/0471140856.tx0112s59 -
BMC Developmental Biology Apr 2014Mosquito eggshells show remarkable diversity in physical properties and structure consistent with adaptations to the wide variety of environments exploited by these...
BACKGROUND
Mosquito eggshells show remarkable diversity in physical properties and structure consistent with adaptations to the wide variety of environments exploited by these insects. We applied proteomic, transcriptomic, and hybridization in situ techniques to identify gene products and pathways that participate in the assembly of the Aedes aegypti eggshell. Aedes aegypti population density is low during cold and dry seasons and increases immediately after rainfall. The survival of embryos through unfavorable periods is a key factor in the persistence of their populations. The work described here supports integrated vector control approaches that target eggshell formation and result in Ae. aegypti drought-intolerant phenotypes for public health initiatives directed to reduce mosquito-borne diseases.
RESULTS
A total of 130 proteins were identified from the combined mass spectrometric analyses of eggshell preparations.
CONCLUSIONS
Classification of proteins according to their known and putative functions revealed the complexity of the eggshell structure. Three novel Ae. aegypti vitelline membrane proteins were discovered. Odorant-binding and cysteine-rich proteins that may be structural components of the eggshell were identified. Enzymes with peroxidase, laccase and phenoloxidase activities also were identified, and their likely involvements in cross-linking reactions that stabilize the eggshell structure are discussed.
Topics: Aedes; Amino Acid Sequence; Animals; Conserved Sequence; Egg Proteins; Electrophoresis, Polyacrylamide Gel; Gene Expression Regulation, Developmental; Genome, Insect; Insect Proteins; Mass Spectrometry; Proteomics; RNA, Messenger
PubMed: 24707823
DOI: 10.1186/1471-213X-14-15 -
PLoS Computational Biology Mar 2014The Drosophila eggshell constitutes a remarkable system for the study of epithelial patterning, both experimentally and through computational modeling. Dorsal eggshell...
The Drosophila eggshell constitutes a remarkable system for the study of epithelial patterning, both experimentally and through computational modeling. Dorsal eggshell appendages arise from specific regions in the anterior follicular epithelium that covers the oocyte: two groups of cells expressing broad (roof cells) bordered by rhomboid expressing cells (floor cells). Despite the large number of genes known to participate in defining these domains and the important modeling efforts put into this developmental system, key patterning events still lack a proper mechanistic understanding and/or genetic basis, and the literature appears to conflict on some crucial points. We tackle these issues with an original, discrete framework that considers single-cell models that are integrated to construct epithelial models. We first build a phenomenological model that reproduces wild type follicular epithelial patterns, confirming EGF and BMP signaling input as sufficient to establish the major features of this patterning system within the anterior domain. Importantly, this simple model predicts an instructive juxtacrine signal linking the roof and floor domains. To explore this prediction, we define a mechanistic model that integrates the combined effects of cellular genetic networks, cell communication and network adjustment through developmental events. Moreover, we focus on the anterior competence region, and postulate that early BMP signaling participates with early EGF signaling in its specification. This model accurately simulates wild type pattern formation and is able to reproduce, with unprecedented level of precision and completeness, various published gain-of-function and loss-of-function experiments, including perturbations of the BMP pathway previously seen as conflicting results. The result is a coherent model built upon rules that may be generalized to other epithelia and developmental systems.
Topics: Animals; Bone Morphogenetic Proteins; Computational Biology; Computer Simulation; Drosophila melanogaster; Egg Proteins; Epidermal Growth Factor; Epithelium; Gene Expression Regulation, Developmental; Gene Regulatory Networks; Morphogenesis; Mutation; Oocytes; Oogenesis; Signal Transduction; Software; Vitelline Membrane
PubMed: 24675973
DOI: 10.1371/journal.pcbi.1003527 -
Fly 2013Drosophila embryo dorsoventral polarity is established by a maternally encoded signal transduction pathway in which three sequentially acting serine proteases,...
Drosophila embryo dorsoventral polarity is established by a maternally encoded signal transduction pathway in which three sequentially acting serine proteases, Gastrulation Defective, Snake and Easter, generate the ligand that activates the Toll receptor on the ventral side of the embryo. The spatial regulation of this pathway depends upon ventrally restricted expression of the Pipe sulfotransferase in the ovarian follicle during egg formation. Several recent observations have advanced our understanding of the mechanism regulating the spatially restricted activation of Toll. First, several protein components of the vitelline membrane layer of the eggshell have been determined to be targets of Pipe-mediated sulfation. Second, the processing of Easter by Snake has been identified as the first Pipe-dependent, ventrally-restricted processing event in the pathway. Finally, Gastrulation Defective has been shown to undergo Pipe-dependent, ventral localization within the perivitelline space and to facilitate Snake-mediated processing of Easter. Together, these observations suggest that Gastrulation Defective, localized on the interior ventral surface of the eggshell in association with Pipe-sulfated eggshell proteins, recruits and mediates an interaction between Snake and Easter. This event leads to ventrally-restricted processing and activation of Easter and consequently, localized formation of the Toll ligand, and Toll activation.
Topics: Animals; Body Patterning; Drosophila; Drosophila Proteins; Embryonic Development; Female; Serine Endopeptidases
PubMed: 24047959
DOI: 10.4161/fly.25141 -
Comptes Rendus Biologies Jul 2013The early intrauterine embryonic development of the bothriocephalidean cestode Clestobothrium crassiceps (Rudolphi, 1819), a parasite of the teleost Merluccius...
Early intrauterine embryonic development of the bothriocephalidean cestode Clestobothrium crassiceps (Rudolphi, 1819), a parasite of the teleost Merluccius merluccius (L., 1758) (Gadiformes: Merlucciidae).
The early intrauterine embryonic development of the bothriocephalidean cestode Clestobothrium crassiceps (Rudolphi, 1819), a parasite of the teleost Merluccius merluccius (L., 1758), was studied by means of light (LM) and transmission electron microscopy (TEM). Contrary to the generic diagnosis given in the CABI Keys to the cestode parasites of vertebrates, the eggs of C. crassiceps, the type of species of Clestobothrium Lühe, 1899, are operculate and embryonated. Our LM and TEM results provide direct evidence that an operculum is present and that the eggs exhibit various stages of intrauterine embryonic development, and in fact represent a good example of early ovoviviparity. The intrauterine eggs of this species are polylecithal and contain numerous vitellocytes, generally ∼30, which are pushed to the periphery and remain close to the eggshell, whereas the dividing zygote and later the early embryo remain in the egg centre. During early intrauterine embryonic development, several cleavage divisions take place, which result in the formation of three types of blastomeres, i.e. macro-, meso- and micromeres. These can be readily differentiated at the TEM level, not only by their size, but also by the ultrastructural characteristics of their nuclei and cytoplasmic organelles. The total number of blastomeres in these early embryos, enclosed within the electron-dense eggshells, can be up to ∼20 cells of various sizes and characteristics. Mitotic divisions of early blastomeres were frequently observed at both LM and TEM levels. Simultaneously with the mitotic cleavage divisions leading to blastomere multiplication and their rapid differentiation, there is also a deterioration of some blastomeres, mainly micromeres. A similar degeneration of vitellocytes begins even earlier. Both processes show a progressive degeneration of both vitellocytes and micromeres, and are good examples of apoptosis, a process that provides nutritive substances, including lipids, for the developing embryo.
Topics: Animals; Blastomeres; Cell Nucleus; Cestoda; Cytoplasm; Embryo, Nonmammalian; Embryonic Development; Female; Freezing; Gadiformes; Microscopy, Electron, Transmission; Mitosis; Oviparity; Parasites; Uterus; Vitelline Membrane
PubMed: 23932252
DOI: 10.1016/j.crvi.2013.06.002 -
International Braz J Urol : Official... 2013To screen proteins/peptides in urine of Renal Cell Carcinoma (RCC) patients by SELDI-TOF (Surface Enhanced Laser Desorption Ionization - Time of Flight) in search of...
PURPOSE
To screen proteins/peptides in urine of Renal Cell Carcinoma (RCC) patients by SELDI-TOF (Surface Enhanced Laser Desorption Ionization - Time of Flight) in search of possible biomarkers.
MATERIAL AND METHODS
Sixty-one urines samples from Clear Cell RCC and Papillary RCC were compared to 29 samples of control urine on CM10 chip. Mass analysis was performed in a ProteinChip Reader PCS 4,000 (Ciphergen Biosystems, Fremont, CA) with the software Ciphergen Express 3.0. All chips were read at low and at high laser energy. For statistical analysis the urine samples were clustered according to the histological classification (Clear Cell and Papillary Carcinoma). For identification urine was loaded on a SDS PAGE gel and bands of most interest were excised, trypsinized and identified by MS/MS. Databank searches were performed in Swiss-Prot database using the MASCOT search algorithm and in Profound.
RESULTS
Proteins that were identified from urine of controls included immunoglobulin light chains, albumin, secreted and transmembrane 1 precursor (protein K12), mannan-binding lectin-associated serine protease-2 (MASP-2) and vitelline membrane outer layer 1 isoform 1. Identification of immunoglobulins and isoforms of albumin are quite common by proteomics and therefore cannot be considered as possible molecular markers. K12 and MASP-2 play important physiological roles, while vitellite membrane outer layer 1 role is unknown since it was never purified in humans.
CONCLUSIONS
The down expression of Protein K-12 and MASP-2 make them good candidates for RCC urine marker and should be validated in a bigger cohort including the other less common histological RCC subtypes.
Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Brazil; Carcinoma, Renal Cell; Cohort Studies; Female; Humans; Kidney Neoplasms; Male; Middle Aged; Neoplasm Staging; Proteomics; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 23683669
DOI: 10.1590/S1677-5538.IBJU.2013.02.12 -
G3 (Bethesda, Md.) May 2013Mating and consequent reproduction significantly reduce the ability of female Drosophila melanogaster to defend against systemic bacterial infection. The goal of the...
Mating and consequent reproduction significantly reduce the ability of female Drosophila melanogaster to defend against systemic bacterial infection. The goal of the present study was to identify genes likely to inform the mechanism of this post-mating immunosuppression. We used microarrays to contrast genome-wide transcript levels in virgin vs. mated females before and after infection. Because the immunosuppressive effect of mating is contingent on the presence of a germline in females, we repeated the entire experiment by using female mutants that do not form a germline. We found that multiple genes involved in egg production show reduced expression in response to infection, and that this reduction is stronger in virgins than it is in mated females. In germline-less females, expression of egg-production genes was predictably low and not differentially affected by infection. We also identified several immune responsive genes that are differentially induced after infection in virgins vs. mated females. Immune genes affected by mating status and egg production genes altered by infection are candidates to inform the mechanism of the trade-off between mating and immune defense.
Topics: Animals; Drosophila melanogaster; Enterobacteriaceae Infections; Female; Gene Expression Regulation; Genes, Insect; Male; Ovum; Providencia; RNA, Messenger; Reproduction; Transcriptome; Vitelline Membrane
PubMed: 23550122
DOI: 10.1534/g3.112.005306 -
FEBS Letters Apr 2013Previous studies have shown that the 3' UTR of BmVMP23 was destroyed by an inserted fragment, and the BmVMP23 expression was downregulated in the lethal egg of "Ming"...
Previous studies have shown that the 3' UTR of BmVMP23 was destroyed by an inserted fragment, and the BmVMP23 expression was downregulated in the lethal egg of "Ming" (l-e(m)). In this study, we found a miRNA (bmo-miR-1a-3p) that matches the 3' UTR sequence of BmVMP23 perfectly. The relationship between bmo-miR-1a-3p and the BmVMP23 expression was examined by the co-transfection in vitro. The luciferase assay result showed that luc expression was strongly repressed. This result inferred that bmo-miR-1a-3p may downregulate BmVMP23 expression via complementary interaction with the target sites at the 3' UTR.
Topics: 3' Untranslated Regions; Animals; Base Sequence; Bombyx; Cell Line; Egg Proteins; Female; Gene Expression Regulation; Green Fluorescent Proteins; Insect Proteins; Luciferases; MicroRNAs; Microscopy, Fluorescence; Molecular Sequence Data; Mutation; Reverse Transcriptase Polymerase Chain Reaction
PubMed: 23481103
DOI: 10.1016/j.febslet.2013.02.030 -
Evolution & Development Jan 2013Cell surface changes in an egg at fertilization are essential to begin development and for protecting the zygote. Most fertilized eggs construct a barrier around...
Cell surface changes in an egg at fertilization are essential to begin development and for protecting the zygote. Most fertilized eggs construct a barrier around themselves by modifying their original extracellular matrix. This construction usually results from calcium-induced exocytosis of cortical granules, the contents of which in sea urchins function to form the fertilization envelope (FE), an extracellular matrix of cortical granule contents built upon a vitelline layer scaffold. Here, we examined the molecular mechanism of this process in sea stars, a close relative of the sea urchins, and analyze the evolutionary changes that likely occurred in the functionality of this structure between these two organisms. We find that the FE of sea stars is more permeable than in sea urchins, allowing diffusion of molecules in excess of 2 megadaltons. Through a proteomic and transcriptomic approach, we find that most, but not all, of the proteins present in the sea urchin envelope are present in sea stars, including SFE9, proteoliaisin, and rendezvin. The mRNAs encoding these FE proteins accumulated most densely in early oocytes, and then beginning with vitellogenesis, these mRNAs decreased in abundance to levels nearly undetectable in eggs. Antibodies to the SFE9 protein of sea stars showed that the cortical granules in sea star also accumulated most significantly in early oocytes, but different from sea urchins, they translocated to the cortex of the oocytes well before meiotic initiation. These results suggest that the preparation for cell surface changes in sea urchins has been shifted to later in oogenesis, and perhaps reflects the meiotic differences among the species-sea star oocytes are stored in prophase of meiosis and fertilized during the meiotic divisions, as in most animals, whereas sea urchins are one of the few taxons in which eggs have completed meiosis prior to fertilization.
Topics: Animals; Cell Membrane; Cell Membrane Permeability; Developmental Biology; Echinodermata; Extracellular Matrix; Fertilization; Gene Expression Regulation, Developmental; In Situ Hybridization; Mass Spectrometry; Meiosis; Oocytes; Oogenesis; Phylogeny; RNA, Messenger; Sea Urchins; Species Specificity; Zygote
PubMed: 23331915
DOI: 10.1111/ede.12012