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1.
Phys Rev E ; 109(6-2): 065201, 2024 Jun.
Article in English | MEDLINE | ID: mdl-39020911

ABSTRACT

Shock-driven implosions with 100% deuterium (D_{2}) gas fill compared to implosions with 50:50 nitrogen-deuterium (N_{2}D_{2}) gas fill have been performed at the OMEGA laser facility to test the impact of the added mid-Z fill gas on implosion performance. Ion temperature (T_{ion}) as inferred from the width of measured DD-neutron spectra is seen to be 34%±6% higher for the N_{2}D_{2} implosions than for the D_{2}-only case, while the DD-neutron yield from the D_{2}-only implosion is 7.2±0.5 times higher than from the N_{2}D_{2} gas fill. The T_{ion} enhancement for N_{2}D_{2} is observed in spite of the higher Z, which might be expected to lead to higher radiative loss, and higher shock strength for the D_{2}-only versus N_{2}D_{2} implosions due to lower mass, and is understood in terms of increased shock heating of N compared to D, heat transfer from N to D prior to burn, and limited amount of ion-electron-equilibration-mediated additional radiative loss due to the added higher-Z material. This picture is supported by interspecies equilibration timescales for these implosions, constrained by experimental observables. The one-dimensional (1D) kinetic Vlasov-Fokker-Planck code ifp and the radiation hydrodynamic simulation codes hyades (1D) and xrage [1D, two-dimensional (2D)] are brought to bear to understand the observed yield ratio. Comparing measurements and simulations, the yield loss in the N_{2}D_{2} implosions relative to the pure D_{2}-fill implosion is determined to result from the reduced amount of D_{2} in the fill (fourfold effect on yield) combined with a lower fraction of the D_{2} fuel being hot enough to burn in the N_{2}D_{2} case. The experimental yield and T_{ion} ratio observations are relatively well matched by the kinetic simulations, which suggest interspecies diffusion is responsible for the lower fraction of hot D_{2} in the N_{2}D_{2} relative to the D_{2}-only case. The simulated absolute yields are higher than measured; a comparison of 1D versus 2D xrage simulations suggest that this can be explained by dimensional effects. The hydrodynamic simulations suggest that radiative losses primarily impact the implosion edges, with ion-electron equilibration times being too long in the implosion cores. The observations of increased T_{ion} and limited additional yield loss (on top of the fourfold expected from the difference in D content) for the N_{2}D_{2} versus D_{2}-only fill suggest it is feasible to develop the platform for studying CNO-cycle-relevant nuclear reactions in a plasma environment.

2.
Rev Sci Instrum ; 94(4)2023 Apr 01.
Article in English | MEDLINE | ID: mdl-38081238

ABSTRACT

The performance of modern laser-driven inertial confinement fusion (ICF) experiments is degraded by contamination of the deuterium-tritium (DT) fuel with high-Z material during compression. Simulations suggest that this mix can be described by the ion temperature distribution of the implosion, given that such contaminants deviate in temperature from the surrounding DT plasma. However, existing neutron time-of-flight (nTOF) diagnostics only measure the spatially integrated ion temperature. This paper describes the techniques and forward modeling used to develop a novel diagnostic imaging system to measure the spatially resolved ion temperature of an ICF implosion for the first time. The technique combines methods in neutron imaging and nTOF diagnostics to measure the ion temperature along one spatial dimension at yields currently achievable on the OMEGA laser. A detailed forward model of the source and imaging system was developed to guide instrument design. The model leverages neutron imaging reconstruction algorithms, radiation hydrodynamics and Monte Carlo simulations, optical ray tracing, and more. The results of the forward model agree with the data collected on OMEGA using the completed diagnostic. The analysis of the experimental data is still ongoing and will be discussed in a separate publication.

3.
Rev Sci Instrum ; 93(11): 113510, 2022 Nov 01.
Article in English | MEDLINE | ID: mdl-36461553

ABSTRACT

A mix of contaminant mass is a known, performance-limiting factor for laser-driven inertial confinement fusion (ICF). It has also recently been shown that the contaminant mass is not necessarily in thermal equilibrium with the deuterium-tritium plasma [B. M. Haines et al., Nat. Commun. 11, 544 (2020)]. Contaminant mass temperature is one of the dominant uncertainties in contaminant mass estimates. The MixIT diagnostic is a new and potentially transformative diagnostic, capable of spatially resolving ion temperature. The approach combines principles of neutron time-of-flight and neutron imaging diagnostics. The information from the MixIT diagnostic can be used to optimize ICF target and laser drive designs as well as provide key constraints on ICF radiation-hydrodynamic simulations that are critical to contaminant mass estimates. This work details the design and optimization of the major components of the MixIT diagnostic: the neutron aperture, the neutron detector (scintillator), and the recording system.

4.
Rev Sci Instrum ; 92(3): 033532, 2021 Mar 01.
Article in English | MEDLINE | ID: mdl-33820092

ABSTRACT

Great strides have been made in improving the quality of x-ray radiographs in high energy density plasma experiments, enabled in part by innovations in engineering and manufacturing of integrated circuits and materials. As a consequence, the radiographs of today are filled with a great deal of detail, but few of these features are extracted in a systematic way. Analysis techniques familiar to plasma physicists tend toward brittle 1D lineout or Fourier transform type analyses. The techniques applied to process our data have not kept pace with improvements in the quality of our data. Fortunately, the field of computer vision has a wealth of tools to offer, which have been widely used in industrial imaging and, more recently, adopted in biological imaging. We demonstrate the application of computer vision techniques to the analysis of x-ray radiographs from high energy density plasma experiments, as well as give a brief tutorial on the computer vision techniques themselves. These tools robustly extract 2D contours of shocks, boundaries of inhomogeneities, and secondary flows, thereby allowing for increased automation of analysis, as well as direct and quantitative comparisons with simulations.

5.
Philos Trans A Math Phys Eng Sci ; 379(2189): 20200005, 2021 Jan 25.
Article in English | MEDLINE | ID: mdl-33280565

ABSTRACT

A European consortium of 15 laboratories across nine nations have worked together under the EUROFusion Enabling Research grants for the past decade with three principle objectives. These are: (a) investigating obstacles to ignition on megaJoule-class laser facilities; (b) investigating novel alternative approaches to ignition, including basic studies for fast ignition (both electron and ion-driven), auxiliary heating, shock ignition, etc.; and (c) developing technologies that will be required in the future for a fusion reactor. A brief overview of these activities, presented here, along with new calculations relates the concept of auxiliary heating of inertial fusion targets, and provides possible future directions of research and development for the updated European Roadmap that is due at the end of 2020. This article is part of a discussion meeting issue 'Prospects for high gain inertial fusion energy (part 2)'.

6.
Philos Trans A Math Phys Eng Sci ; 379(2189): 20200224, 2021 Jan 25.
Article in English | MEDLINE | ID: mdl-33280567

ABSTRACT

Indirect drive inertial confinement fusion experiments with convergence ratios below 17 have been previously shown to be less susceptible to Rayleigh-Taylor hydrodynamic instabilities, making this regime highly interesting for fusion science. Additional limitations imposed on the implosion velocity, in-flight aspect ratio and applied laser power aim to further reduce instability growth, resulting in a new regime where performance can be well represented by one-dimensional (1D) hydrodynamic simulations. A simulation campaign was performed using the 1D radiation-hydrodynamics code HYADES to investigate the performance that could be achieved using direct-drive implosions of liquid layer capsules, over a range of relevant energies. Results include potential gains of 0.19 on LMJ-scale systems and 0.75 on NIF-scale systems, and a reactor-level gain of 54 for an 8.5 MJ implosion. While the use of 1D simulations limits the accuracy of these results, they indicate a sufficiently high level of performance to warrant further investigations and verification of this new low-instability regime. This potentially suggests an attractive new approach to fusion energy. This article is part of a discussion meeting issue 'Prospects for high gain inertial fusion energy (part 2)'.

7.
Phys Rev E ; 102(4-1): 043212, 2020 Oct.
Article in English | MEDLINE | ID: mdl-33212701

ABSTRACT

In this work, we present results from experiments capable of producing and measuring the propagation of multiple successive, copropagating shocks across an unstable planar interface, where the shocks are independently driven and separately controllable, enabling the study of this important phenomenon. Copropagating shocks play a significant role in a wide range of systems involving stratified media subject to a shock, and exhibit different physical characteristics compared to counterpropagating shocks. Existing techniques, however, preclude copropagating shocks, so experiments to date have been limited to the study of counterpropagating shocks. We address this previous limitation and open a physical parameter space for study using a new hohlraum platform on the National Ignition Facility. Initial experimental results are presented together with comparisons from numerical simulations.

8.
NPJ Genom Med ; 4: 27, 2019.
Article in English | MEDLINE | ID: mdl-31700678

ABSTRACT

A growing body of evidence points to a considerable and heterogeneous genetic aetiology of cerebral palsy (CP). To identify recurrently variant CP genes, we designed a custom gene panel of 112 candidate genes. We tested 366 clinically unselected singleton cases with CP, including 271 cases not previously examined using next-generation sequencing technologies. Overall, 5.2% of the naïve cases (14/271) harboured a genetic variant of clinical significance in a known disease gene, with a further 4.8% of individuals (13/271) having a variant in a candidate gene classified as intolerant to variation. In the aggregate cohort of individuals from this study and our previous genomic investigations, six recurrently hit genes contributed at least 4% of disease burden to CP: COL4A1, TUBA1A, AGAP1, L1CAM, MAOB and KIF1A. Significance of Rare VAriants (SORVA) burden analysis identified four genes with a genome-wide significant burden of variants, AGAP1, ERLIN1, ZDHHC9 and PROC, of which we functionally assessed AGAP1 using a zebrafish model. Our investigations reinforce that CP is a heterogeneous neurodevelopmental disorder with known as well as novel genetic determinants.

9.
Phys Rev Lett ; 118(13): 135001, 2017 Mar 31.
Article in English | MEDLINE | ID: mdl-28409959

ABSTRACT

We present narrow-band self-emission x-ray images from a titanium tracer layer placed at the fuel-shell interface in 60-laser-beam implosion experiments at the OMEGA facility. The images are acquired during deceleration with inferred convergences of ∼9-14. Novel here is that a systematically observed asymmetry of the emission is linked, using full sphere 3D implosion modeling, to performance-limiting low mode asymmetry of the drive.

10.
Phys Rev Lett ; 117(24): 245001, 2016 Dec 09.
Article in English | MEDLINE | ID: mdl-28009190

ABSTRACT

The first cryogenic deuterium and deuterium-tritium liquid layer implosions at the National Ignition Facility (NIF) demonstrate D_{2} and DT layer inertial confinement fusion (ICF) implosions that can access a low-to-moderate hot-spot convergence ratio (1230) DT ice layer implosions. Although high CR is desirable in an idealized 1D sense, it amplifies the deleterious effects of asymmetries. To date, these asymmetries prevented the achievement of ignition at the NIF and are the major cause of simulation-experiment disagreement. In the initial liquid layer experiments, high neutron yields were achieved with CRs of 12-17, and the hot-spot formation is well understood, demonstrated by a good agreement between the experimental data and the radiation hydrodynamic simulations. These initial experiments open a new NIF experimental capability that provides an opportunity to explore the relationship between hot-spot convergence ratio and the robustness of hot-spot formation during ICF implosions.

11.
New Phytol ; 155(3): 363-372, 2002 Sep.
Article in English | MEDLINE | ID: mdl-33873323

ABSTRACT

• Thlaspi caerulescens hyper-accumulates heavy metals, and localized root proliferation in response to patchily distributed metals may enhance heavy metal uptake. Here, an experimental assessment is made of the impact of heterogeneous zinc (Zn) supply, and of the contrast between the concentration of Zn in soil patches, on Zn hyperaccumulation, patterns of root placement, growth, biomass allocation patterns and nutrient acquisition in two ecotypes of T. caerulescens . • Growth of Prayon and Bradford Dale ecotypes (which originate from a heavily contaminated and a less Zn-contaminated site, respectively) was compared in treatments with either heterogeneous or homogeneous Zn supply and in a treatment with no Zn added to the substrate. • Prayon exhibited enhanced above- and below-ground biomass production, and zincophilic root foraging patterns in response to heterogeneously distributed Zn. The root placement pattern suggested that T. caerulescens may discriminate between patches with contrasting Zn concentrations and produce more roots in patches with higher Zn concentrations. Bradford Dale did not show these responses. • The T. caerulescens ecotypes may differ in their Zn requirements or may have experienced selection for genetic differences that affect their ability to respond to Zn heterogeneity. Such differences will be important factors in the successful phytoremediation of contaminated soils.

12.
J Immunol ; 167(11): 6644-53, 2001 Dec 01.
Article in English | MEDLINE | ID: mdl-11714836

ABSTRACT

Chemokine receptors are differentially expressed on immature and mature dendritic cells (DC). Herein, we demonstrate for the first time that murine antimicrobial peptides beta-defensins 2 and 3 bind murine CCR6, similarly to inflammatory chemokine macrophage-inflammatory protein 3alpha, and they chemoattract bone marrow-derived immature, but not mature DC. Using various chemokines or defensins fused with nonimmunogenic tumor Ags, we studied their capacity to delivery Ags to subsets of immune cells to elicit antitumor immunity. We demonstrate that DNA immunizations with fusion constructs with beta-defensin 2 or inflammatory chemokines that target immature DC, but not homeostatic chemokines secondary lymphoid tissue chemokine, CCL21, or stromal cell-derived factor 1, CXCL12, which chemoattract mature DC, elicit humoral, protective, and therapeutic immunity against two different syngeneic lymphomas.


Subject(s)
Antigens, Neoplasm/genetics , Antigens, Neoplasm/immunology , Dendritic Cells/cytology , Dendritic Cells/immunology , Immunotherapy, Adoptive/methods , Lymphoma, B-Cell/immunology , Lymphoma, B-Cell/therapy , Vaccines, DNA/immunology , Animals , Antibodies, Anti-Idiotypic/biosynthesis , Antibodies, Anti-Idiotypic/genetics , Cancer Vaccines/administration & dosage , Cancer Vaccines/genetics , Cancer Vaccines/immunology , Cell Differentiation/genetics , Cell Differentiation/immunology , Cell Line , Chemokines/administration & dosage , Chemokines/genetics , Chemokines/immunology , Chemokines/physiology , Female , Gene Targeting , Humans , Immunity, Innate/genetics , Immunoglobulin Variable Region/administration & dosage , Immunoglobulin Variable Region/genetics , Immunoglobulin Variable Region/immunology , Lymphoma, B-Cell/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , Receptors, Chemokine/genetics , Recombinant Fusion Proteins/administration & dosage , Recombinant Fusion Proteins/chemical synthesis , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/physiology , Tumor Cells, Cultured , Vaccines, DNA/administration & dosage , Vaccines, DNA/genetics , beta-Defensins/administration & dosage , beta-Defensins/genetics , beta-Defensins/immunology , beta-Defensins/physiology
13.
J Abnorm Child Psychol ; 29(5): 445-63, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11695545

ABSTRACT

This paper presents an initial assessment of the Children's Attributional Style Interview (CASI), a newly designed measure for assessing attributional style in young children (age 5 and up). The CASI was used to conduct prospective tests of the reformulated helplessness and the integrated hopelessness/self-esteem theories of depression in a sample of 147 5-10-year-old children. For comparison, the same tests were also conducted with the Children's Attributional Style Questionnaire-Revised, a commonly used measure for assessing attributional style in older children (age 8 and up). The CASI evidenced support of the reformulated helplessness theory and partial support of the integrated hopelessness/self-esteem theory. The CASI also demonstrated good internal consistency. Thus, our findings provide initial support for the CASI as a methodologically sound measure of attributional style for children as young as 5 years old. Although preliminary, our findings also suggest possible developmental differences in how attributional style interacts with self-esteem and negative life stress. The CASI should prove to be a useful tool in furthering the understanding of the origins and development of attributional style in childhood, as well as its contribution to the understanding of the development and prevention of depressive symptomatology in children.


Subject(s)
Cognition , Depressive Disorder, Major/diagnosis , Interview, Psychological , Psychological Theory , Social Perception , Child , Child, Preschool , Depressive Disorder, Major/psychology , Humans , Neuropsychological Tests , Prospective Studies , Psychology, Child , Self Concept , Surveys and Questionnaires
14.
J Chem Ecol ; 27(7): 1525-34, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11504042

ABSTRACT

We examined 40 taxa from nine genera within the subfamily Mimosoideae; 29 (73%) from six genera produced carbon disulfide (CS2). In addition, 19 of 40 taxa (48%) produced carbonyl sulfide (OCS). Of nine mimosoid taxa that produced CS2, all possessed a djenkolic acid and a cysteine lyase. Of three mimosoid taxa that did not produce CS2, two lacked a cysteine lyase and one lacked both a lyase and a djenkolic acid. Of 16 taxa from 14 genera from the other two subfamilies of the Fabaceae, the Caesalpinioideae and Papilionoideae, none produced CS2. The results suggest that CS2 production is common in the Mimosoideae and uncommon in the Caesalpinioideae and Papilionoideae and that plants in the Mimosoideae that do produce CS2 must possess both a djenkolic acid and a cysteine lyase for this production to occur.


Subject(s)
Carbon Disulfide/chemistry , Cysteine/analogs & derivatives , Fabaceae/chemistry , Carbon Disulfide/analysis , Carbon-Sulfur Lyases/chemistry , Carbon-Sulfur Lyases/metabolism , Cysteine/chemistry , Cysteine/metabolism , Plant Roots/chemistry , Volatilization
15.
Mol Immunol ; 38(1): 9-18, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11483206

ABSTRACT

Although the mouse immunoglobulin heavy chain (Igh) locus contains 15 heavy chain V (Vh) gene families, at least half of the Vh gene segments are members of the VhJ558 family. This large Vh gene family represents the least characterized germline coding regions of any of the mouse antigen receptor loci and the contribution of individual VhJ558 genes to the preimmune repertoire is poorly understood. In fact, relatively few germline VhJ558 sequences have been reported for BALB/c, the foundation strain for mouse immunoglobulin genetics and the prototypic strain of the Igh(a) haplotype. Here we present a database consisting of 66 sequences estimated to represent one-half of the total number of functional BALB/c VhJ558 genes. Our results indicate that a subset of the VhJ558 genes is highly expressed in the preimmune repertoire, with just nine Vh sequences accounting for nearly 50% of the VhJ558 heavy chains expressed by splenic B cells. We show that this disparity in the expressed Vh gene repertoire is not due to the position of the Vh genes relative to the Dh cluster or to multiple germline copies of the highly expressed VhJ558 genes. Together, these data constitute the first detailed analysis of functional BALB/c VhJ558 genes, demonstrate a striking bias in the use of particular VhJ558 genes in the preimmune repertoire, and provide sufficient information to study the regulation of the Dh-distal region of the Igh-V locus at the level of individual genes.


Subject(s)
Genetic Variation , Germ-Line Mutation , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Variable Region/genetics , Amino Acid Sequence , Animals , Base Sequence , Chromosome Mapping , DNA, Complementary , Gene Expression , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Molecular Sequence Data
16.
Oper Dent ; 26(1): 44-7, 2001.
Article in English | MEDLINE | ID: mdl-11203776

ABSTRACT

This study compared the shear bond strength of four filled dentin bonding agents (Prime & Bond NT, One Coat Bond, OptiBond Solo, PermaQuik1) to a conventional unfilled Dentin Bonding agent (3M Scotchbond Multipurpose Plus). A column of composite resin (Herculite XRV Restorative Resin) was bonded to human dentin using the five dentin-bonding systems following the manufacturers' directions. The specimens were thermocycled 1000 cycles from 6 to 60 degrees C. Each specimen was tested on an MTS machine in shear-to-failure. The data were subjected to One-Factor ANOVA and Newman-Kuels post-hoc tests. Statistical analysis revealed that One Coat Bond had significantly greater shear bond strength than all other materials tested.


Subject(s)
Dental Bonding , Dentin-Bonding Agents/chemistry , Analysis of Variance , Bisphenol A-Glycidyl Methacrylate/chemistry , Composite Resins/chemistry , Dental Stress Analysis/instrumentation , Dentin/ultrastructure , Humans , Materials Testing , Polymethacrylic Acids/chemistry , Resin Cements/chemistry , Statistics as Topic , Stress, Mechanical , Thermodynamics
17.
Biochem J ; 351 Pt 3: 557-65, 2000 Nov 01.
Article in English | MEDLINE | ID: mdl-11042109

ABSTRACT

Nedd4 is a member of a growing family of ubiquitin-protein ligases which consist of a lipid-binding domain, two to four WW domains and a C-terminal ubiquitin-protein ligase domain. The Nedd4 mRNA levels are developmentally regulated and Nedd4 protein is highly expressed in many mouse embryonic tissues. In this study we have used a far-Western screen to identify embryonic proteins that interact with the WW domains in mouse Nedd4. We report here identification of eight Nedd4 WW-domain-interacting proteins from mouse embryonic cDNA expression libraries. Two of the proteins are novel, while two have been identified previously as ligands for a WW domain. All of these proteins contain one or more PY motifs. In seven of the eight proteins, these PY motifs are necessary for their interaction with the WW domains of Nedd4. Using site-directed mutagenesis, and by using individual WW domains of Nedd4 as probes for far-Western analysis, we show that the three WW domains in Nedd4 interact with varying affinities with the PY motifs present in various Nedd4-binding proteins. These results provide evidence that Nedd4 can potentially interact with multiple proteins, possibly simultaneously, through its WW domains.


Subject(s)
Calcium-Binding Proteins/metabolism , Embryo, Mammalian/metabolism , Ligases/metabolism , Ubiquitin-Protein Ligases , Amino Acid Motifs , Amino Acid Sequence , Animals , Calcium-Binding Proteins/chemistry , Calcium-Binding Proteins/genetics , Endosomal Sorting Complexes Required for Transport , Gene Expression Regulation, Developmental , Humans , Ligases/chemistry , Ligases/genetics , Membrane Proteins/chemistry , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mice , Molecular Sequence Data , Nedd4 Ubiquitin Protein Ligases , Protein Binding , RNA, Messenger/genetics
18.
Mol Biol Cell ; 11(4): 1369-83, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10749936

ABSTRACT

Although many growth factors and cytokines have been shown to be localized within the cell and nucleus, the mechanism by which these molecules elicit a biological response is not well understood. The cytokine leukemia inhibitory factor (LIF) provides a tractable experimental system to investigate this problem, because translation of alternatively spliced transcripts results in the production of differentially localized LIF proteins, one secreted from the cell and acting via cell surface receptors and the other localized within the cell. We have used overexpression analysis to demonstrate that extracellular and intracellular LIF proteins can have distinct cellular activities. Intracellular LIF protein is localized to both nucleus and cytoplasm and when overexpressed induces apoptosis that is inhibited by CrmA but not Bcl-2 expression. Mutational analysis revealed that the intracellular activity was independent of receptor interaction and activation and reliant on a conserved leucine-rich motif that was not required for activation of cell surface receptors by extracellular protein. This provides the first report of alternate intracellular and extracellular cytokine activities that result from differential cellular localization of the protein and are mediated by spatially distinct motifs.


Subject(s)
Growth Inhibitors/metabolism , Interleukin-6 , Lymphokines/metabolism , Viral Proteins , Amino Acid Motifs , Amino Acid Sequence , Animals , Apoptosis/physiology , COS Cells , Cell Line , Cell Nucleus/metabolism , Cytoplasm/metabolism , Extracellular Space/metabolism , Fluorescent Antibody Technique , Growth Inhibitors/chemistry , Leucine/chemistry , Leukemia Inhibitory Factor , Lymphokines/chemistry , Molecular Sequence Data , Proto-Oncogene Proteins c-bcl-2/metabolism , Receptors, Cell Surface/physiology , Sequence Analysis, Protein , Serpins/metabolism , Transfection
19.
Zygote ; 7(3): 239-48, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10533707

ABSTRACT

The zona pellucida (ZP) is an extracellular glycoprotein coat that is deposited around the oocyte during folliculogenesis and performs several functions that relate to fertilisation and preimplantion development. In eutherian mammals it consists of three major glycoproteins--ZPA, ZPB, and ZPC--but little is known about its molecular constitution in marsupials. We have isolated the cDNA encoding the ZPA homologue in two distantly related marsupial series: the possum, Trichosurus vulpecula (a phalangerid) and the dunnart Sminthopsis crassicaudata (a dasyurid). The two cDNA sequences were 86% identical and showed extensive regions of homology to eutherian ZPA proteins, particularly in the central region of the molecule. Many other features of the ZPA protein, except the positioning of the N-linked glycosylation sites, were also conserved between marsupials and eutherians. ZPA expression was shown to occur maximally in the cytoplasm of the oocyte primary follicles with a little, but significant, expression in oocytes of both primordial follicles and in the cytoplasm of the oocyte in follicles with an antral cavity. No expression was seen in surrounding follicle or granulosa cells.


Subject(s)
Egg Proteins/metabolism , Marsupialia/genetics , Membrane Glycoproteins/metabolism , Oocytes/metabolism , Receptors, Cell Surface , Zona Pellucida/metabolism , Amino Acid Sequence , Animals , Base Sequence , Egg Proteins/genetics , Egg Proteins/isolation & purification , Female , In Situ Hybridization , Membrane Glycoproteins/genetics , Membrane Glycoproteins/isolation & purification , Molecular Sequence Data , Ovarian Follicle/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Transcription, Genetic , Zona Pellucida Glycoproteins
20.
Exp Cell Res ; 249(2): 199-211, 1999 Jun 15.
Article in English | MEDLINE | ID: mdl-10366419

ABSTRACT

The polyfunctional cytokine leukemia inhibitory factor (LIF) has been implicated in the maintenance of many stem and progenitor cell populations and as an autocrine growth factor for many tumor cell populations, including germ cell tumors. Studies of LIF transcript expression in germ cell tumor cell lines identified two novel human LIF transcripts, hLIF-M and hLIF-T, containing noncoding alternate first exons that are conserved among all reported LIF genes. Embryonal carcinoma (EC) cell lines expressed these transcripts at consistent levels and hLIF-M was generally the predominant LIF transcript in these cells. This expression pattern was characteristic of EC cells since variable independently regulated expression of these transcripts was evident in other cell lines. Overexpression analysis demonstrated that each alternate hLIF transcript generated different levels of extracellular LIF activity as a consequence of the translation of distinct but partially overlapping sets of proteins. Secreted LIF proteins translated from alternate initiation codons were expressed from the hLIF-D and hLIF-M transcripts. Intracellular, potentially cell-autonomous, proteins were encoded by the hLIF-M and hLIF-T transcripts. Since EC cell lines also expressed LIF receptor transcripts, the novel LIF transcription profiles and proteins identified here suggest a role for autocrine and/or cell-autonomous LIF signaling during germ cell tumorigenesis.


Subject(s)
Carcinoma, Embryonal/metabolism , Germinoma/metabolism , Growth Inhibitors/biosynthesis , Growth Inhibitors/genetics , Interleukin-6 , Lymphokines/biosynthesis , Lymphokines/genetics , Alternative Splicing/drug effects , Base Sequence , Carcinoma, Embryonal/genetics , Cell Differentiation/drug effects , Cell Differentiation/genetics , Cells, Cultured , Exons , Extracellular Space/metabolism , Gene Expression Regulation/drug effects , Genes, Overlapping , Germinoma/genetics , Growth Inhibitors/metabolism , Humans , Leukemia Inhibitory Factor , Lymphokines/metabolism , Molecular Sequence Data , Protein Isoforms/genetics , Protein Isoforms/metabolism , Transcription, Genetic/drug effects , Tretinoin/pharmacology , Tumor Cells, Cultured
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