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Evolutionary origin and phylogenetic analysis of the novel oocyte-specific eukaryotic translation initiation factor 4E in Tetrapoda.
ABSTRACT The transcriptionally active, growing oocyte accumulates mRNAs essential for early stages of development, the oocyte-to-embryo transition, in a stable, dormant form. Translational repression of mRNAs in eggs of various species is conferred by interactions, either direct or via intermediate proteins, of repressive factors bound to the 3'-untranslated regions with the proteins of the eukaryotic translation initiation factor 4E (eIF4E) family bound to the 5'-cap of the transcripts. Recently, a novel oocyte-specific eIF4E encoded by the Eif41b gene in mammals has been identified by our group. To further investigate this gene, the available cDNA libraries, as well as genome assemblies of nonmammalian vertebrates, were surveyed. This analysis revealed that the Eif4e1b gene arose in Tetrapoda as a result of the ancestral Eif4e locus duplication. Unlike other known proteins of three subfamilies comprising eIF4E family (eIF4E1, eIF4E2, and eIF4E3), cDNA library evidence suggests that Eif41b locus has an oocyte-restricted expression across all classes of Tetrapoda. To further understand the role of eIF4E1B during oocyte maturation, injections of antisense morpholino nucleotides in the X. tropicalis fully-grown stage VI oocytes were performed. The resulted ablation of eIF4E1B protein led to significant acceleration of oocyte maturation after progesterone induction; morpholino-injected oocytes formed the metaphase plate 30 min faster than the control groups. These results suggest that eIF4E1B protein acts as a repressor in translational regulation of maternal mRNAs activated during, and required for, oocyte maturation.
- SourceAvailable from: ncbi.nlm.nih.gov
Article: KHDC1B is a novel CPEB binding partner specifically expressed in mouse oocytes and early embryos.[Show abstract] [Hide abstract]
ABSTRACT: mRNAs required for meiotic maturation and early embryonic development are stored in growing oocytes. These transcripts are translationally repressed until hormonal cues trigger ovulation. Errors in translation underlie some cases of human infertility and are associated with ovarian germ cell tumors. However, it remains unclear how maternal transcripts are kept quiescent in mammals. This study describes a potential translational regulator, KHDC1B. KHDC1B is a member of a small family of KH-domain containing proteins specific to eutherian mammals. Two family members, KHDC1A and 1B, are highly expressed in oocytes. KHDC1A and 1B bind polyU agarose and form oligomers like other KH-domain proteins. The functions of these proteins were tested by expression in Xenopus embryos. KHDC1A caused cell death, whereas KHDC1B caused cleavage arrest. This arrest phenotype was rescued by coexpression of the mouse translational regulator cytoplasmic polyadenylation binding protein 1 (mCPEB1). Coimmunoprecipitation and coimmunostaining experiments confirmed the functional interaction between KHDC1B and mCPEB1. Finally, KHDC1B levels and binding partners were shown to fluctuate with the cell cycle. KHDC1B, via its interaction with mCEPB1, may regulate translation of mRNA targets required for oocyte maturation.Molecular biology of the cell 09/2010; 21(18):3137-48. · 5.98 Impact Factor
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ABSTRACT: Formation of the germline in an embryo marks a fresh round of reproductive potential, yet the developmental stage and location within the embryo where the primordial germ cells (PGCs) form differs wildly among species. In most animals, the germline is formed either by an inherited mechanism, in which maternal provisions within the oocyte drive localized germ-cell fate once acquired in the embryo, or an inductive mechanism that involves signaling between cells that directs germ-cell fate. The inherited mechanism has been widely studied in model organisms such as Drosophila melanogaster, Caenorhabditis elegans, Xenopus laevis, and Danio rerio. Given the rapid generation time and the effective adaptation for laboratory research of these organisms, it is not coincidental that research on these organisms has led the field in elucidating mechanisms for germline specification. The inductive mechanism, however, is less well understood and is studied primarily in the mouse (Mus musculus). In this review, we compare and contrast these two fundamental mechanisms for germline determination, beginning with the key molecular determinants that play a role in the formation of germ cells across all animal taxa. We next explore the current understanding of the inductive mechanism of germ-cell determination in mice, and evaluate the hypotheses for selective pressures on these contrasting mechanisms. We then discuss the hypothesis that the transition between these determination mechanisms, which has happened many times in phylogeny, is more of a continuum than a binary change. Finally, we propose an analogy between germline determination and sex determination in vertebrates-two of the milestones of reproduction and development-in which animals use contrasting strategies to activate similar pathways. Mol. Reprod. Dev. © 2013 Wiley Periodicals, Inc.Molecular Reproduction and Development 02/2013; · 2.81 Impact Factor
Article: A C. elegans eIF4E-family member upregulates translation at elevated temperatures of mRNAs encoding MSH-5 and other meiotic crossover proteins.[Show abstract] [Hide abstract]
ABSTRACT: Caenorhabditis elegans expresses five family members of the translation initiation factor eIF4E whose individual physiological roles are only partially understood. We report a specific role for IFE-2 in a conserved temperature-sensitive meiotic process. ife-2 deletion mutants have severe temperature-sensitive chromosome-segregation defects. Mutant germ cells contain the normal six bivalents at diakinesis at 20 degrees C but 12 univalents at 25 degrees C, indicating a defect in crossover formation. Analysis of chromosome pairing in ife-2 mutants at the permissive and restrictive temperatures reveals no defects. The presence of RAD-51-marked early recombination intermediates and 12 well condensed univalents indicate that IFE-2 is not essential for formation of meiotic double-strand breaks or their repair through homologous recombination but is required for crossover formation. However, RAD-51 foci in ife-2 mutants persist into inappropriately late stages of meiotic prophase at 25 degrees C, similar to mutants defective in MSH-4/HIM-14 and MSH-5, which stabilize a critical intermediate in crossover formation. In wild-type worms, mRNAs for msh-4/him-14 and msh-5 shift from free messenger ribonucleoproteins to polysomes at 25 degrees C but not in ife-2 mutants, suggesting that IFE-2 translationally upregulates synthesis of MSH-4/HIM-14 and MSH-5 at elevated temperatures to stabilize Holliday junctions. This is confirmed by an IFE-2-dependent increase in MSH-5 protein levels.Journal of Cell Science 07/2010; 123(Pt 13):2228-37. · 5.88 Impact Factor
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Evolutionary origin and phylogenetic analysis of the novel oocyte-
specific eukaryotic translation initiation factor 4E in Tetrapoda.
Alexei V. Evsikov1 and C. Marín de Evsikova
The transcriptionally active, growing oocyte accumulates mRNAs essential for early stages of
development, the oocyte-to-embryo transition, in a stable, dormant form. Translational repression of
mRNAs in eggs of various species is conferred by interactions, either direct or via intermediate
proteins, of repressive factors bound to the 3′-UTRs with the proteins of the eukaryotic translation
initiation factor 4E (eIF4E) family bound to the 5′-cap of the transcripts. Recently, a novel oocyte-
specific eIF4E encoded by the Eif41b gene in mammals has been identified by our group. To further
investigate this gene, the available cDNA libraries, as well as genome assemblies of non-mammalian
vertebrates were surveyed. This analysis revealed that the Eif4e1b gene arose in Tetrapoda as a result
of the ancestral Eif4e locus duplication. Unlike other known proteins of three subfamilies comprising
eIF4E family (eIF4E1, eIF4E2 and eIF4E3), cDNA library evidence suggests that Eif41b locus has
an oocyte-restricted expression across all classes of Tetrapoda. To further understand the role of
eIF4E1B during oocyte maturation, injections of antisense morpholino nucleotides in the X.
tropicalis fully-grown stage VI oocytes were performed. The resulted ablation of eIF4E1B protein
led to significant acceleration of oocyte maturation after progesterone induction; morpholino-injected
oocytes formed the metaphase plate 30 minutes faster than the control groups. These results suggest
that eIF4E1B protein acts as a repressor in translational regulation of maternal mRNAs activated
during, and required for, oocyte maturation.
comparative genomics; eukaryotic translation initiation factor 4E; molecular evolution; oocyte
maturation; translational control
The early development of metazoans relies heavily on the maternal stores accumulated in the
egg during its growth. Amassing of nutrients during oogenesis secures an embryo's
independence from external energy sources and ensures rapid development during this
vulnerable stage of an organism's life. Another vital role played by the egg is imparting the
initial programs for gene expression upon the nascent embryonic genome. This role is
particularly pronounced in viviparous mammals, whose oocytes have the minute amounts of
yolk and embryos receive nutrition directly from mothers upon implantation. During the
ooplasm-driven remodeling of parental genomes, transcription is barely, if at all, detectable.
Thus, the initial embryonic genome is shaped by precise post-transcriptional and post-
translational mechanisms, which concomitantly create the dynamic molecular environment as
early development proceeds. Upon completion of this genome remodeling, re-initiation of
1 Address correspondence to: The Jackson Laboratory 600 Main Street Bar Harbor, ME 04609, USA Phone: +1-207-288-6613 Facsimile:
+1-207-288-6131 firstname.lastname@example.com.
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transcription demarcates 'embryonic genome activation'. Henceforth, the embryonic genome
gradually assumes control of development from 'maternal command'.
The molecules that drive embryogenesis at these earliest stages are still poorly understood.
However, it is well-known that post-translational mechanisms, which govern initial embryonic
development, rely on massive accumulation of transcripts during oogenesis. Indeed, during
oocyte growth, many transcribed mRNAs are stored in the ooplasm as a 'dormant' form for
translation at later stages and are deadenylated. Translational recruitment of these stored
mRNAs is typically manifested by lengthening of their limited 3′ poly(A) tails (Bachvarova,
1992; Richter and Sonenberg, 2005). Spatio-temporal regulation of translational activation of
maternal mRNAs deposited during oocyte growth is a key mechanism providing new proteins
during the final stages of oogenesis (i.e., maturation), and first stages of embryogenesis in
virtually all metazoans. One model for mRNA translational regulation, based upon extensive
studies from diverse model organisms, proposes how transcripts may be translated in the time-
and stage-dependent fashion (Richter and Sonenberg, 2005; Standart and Minshall, 2008). In
this model, a regulatory protein can be bound to a cognate motif in the 3′-untranslated region
(UTR) of mRNA. Concurrently, this RNA-binding protein may also be bound, either directly
or via intermediate protein(s), to the eukaryotic initiation factor 4E (eIF4E), an mRNA 5′-cap-
binding protein. In the 'closed' conformation, these protein-RNA interactions suppress
translation by preventing formation of the pre-initiation complex. Certain signaling events,
such as phosphorylation of a 3′-UTR-bound regulatory protein, result in a conformational
change to the complex, leading to recruitment of poly(A) polymerases to the 3′ end of mRNA,
formation of the translational initiation complex on the 5′ end and eventually, leading to
The eIF4E family of proteins consists of three large subfamilies, eIF4E1, eIF4E2 and eIF4E3.
Genomic sequencing and analysis of various model organisms, as well as the analysis of EST
databases, suggest the existence of multiple eIF4E-encoding genes within each of these
subfamilies in most organisms (Joshi et al., 2005). The reason for this redundancy is not clear.
While it is assumed that most eIF4E proteins possess at least some 5′-cap binding activity, to
date, the most extensively studied and characterized is the eIF4E1 subfamily. Until recently,
only one mammalian eIF4E1 member, encoded by the Eif4e gene, has been identified. Our
analysis of the fully-grown mouse oocyte cDNA library revealed another gene, Eif4eloo
(Eif4e1b in current nomenclature), which, indeed, is a second bona fide member of this
subfamily in mammals (Evsikov et al., 2006). Unlike its ubiquitously expressed paralog
Eif4e, transcription of Eif4e1b gene appears restricted to growing oocytes. To date, the
Eif4e1b gene has been identified in genome assemblies of various mammals (Evsikov et al.,
2006), while transcripts encoding eIF4E1B proteins from different species, including Homo
sapiens, have been identified in the large-scale survey of GenBank mRNAs and ESTs aimed
to categorize all available information on various eIF4E family proteins across all eukaryotes
(Joshi et al., 2005).
In order to understand the role of Eif4e1b/Eif4eloo in the oocytes, we sought to delineate the
phylogenetic history of this gene. Indeed, a similar eIF4E1 homolog, with the expression
pattern restricted to oocytes, testis and muscle, has been reported in the zebrafish, Danio
rerio (Robalino et al., 2004). Interestingly, unlike 'conventional' eIF4E, this D. rerio eIF4E-1B
protein cannot substitute for endogenous eIF4E in complementation assays using eif4e-
deficient Saccharomyces cerevisiae; furthermore, it lacks the 5′-cap-binding activity (Robalino
et al., 2004). Recently, a similar eIF4E1B protein with similar biochemical properties has been
also identified in the frog, Xenopus laevis (Minshall et al., 2007). Here, we used available
genome sequences and whole-genome assemblies of model organisms to scrutinize the
evolutionary relationships among members of the eIF4E1 subfamily in vertebrates and to
identify the non-mammalian orthologs of Eif4e1b. To investigate the functional role of
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Eif4e1b as a regulator of translation during oocyte maturation, antisense morpholino
oligonucleotides designed to suppress Eif4e1b mRNA translation were injected in immature,
fully-grown Xenopus tropicalis oocytes.
Results and Discussion
To determine the potential orthologs of Eif4e1b/Eif4eloo, we have identified, using TBLASTN
program, the ESTs of three vertebrates, Anolis carolinensis, Gallus gallus and Xenopus
tropicalis, which share identity with Mus musculus eIF4E1B protein. The matching ESTs were
next assembled into contigs, and the identities of resulting consensus cDNAs were established
by aligning them, using BLAT, to the ENSEMBL genome assemblies of A. carolinensis, G.
gallus, and X. tropicalis, respectively. As expected, some consensi aligned to the genomic
regions of A. carolinensis, G. gallus, and X. tropicalis syntenic to the mammalian Eif4e gene,
and the orthology was evidenced by colinearity of genes (Metap1 – Eif4e – Tspan5) in the
genomes of all examined species (Fig. 1A). Another group of EST consensi aligned to the
genomic regions of A. carolinensis and X. tropicalis that were clearly syntenic to the M.
musculus locus (Fig. 1B: Sncb – Eif4e1b – Tspan17). Similarly, one G. gallus cDNA aligned
to the syntenic locus downstream of Sncb gene (Fig. 1B: Sncb – Eif4e1b – Unc5a).
Interestingly, Tspan17 locus is apparently absent in this species as Unc5a, located immediately
downstream of Eif4e1b in G. gallus (Fig. 1B), is located downstream of Tspan17 in other
species (not shown). Virtual Northern blotting (i.e., analysis of the EST sources for the consensi
that comprised Eif4e1b cDNAs in G. gallus, X. tropicalis and A. carolinensis), identified that
all X. tropicalis ESTs were from either oocyte, blastula or gastrula cDNA libraries, while G.
gallus and A. carolinensis ESTs were from ovarian cDNA libraries. Due to the limited amounts
of ESTs available for a marsupial, Monodelphis domestica, Eif4e and Eif4e1b loci in these
species were identified using TBLASTN search of genome assembly using M. musculus
eIF4E1B. This search revealed two candidate genes; further examination of adjacent loci
authenticated these candidates as bona fide orthologs of Eif4e and Eif4e1b, respectively (Fig.
1A, 1B). In summary, we have identified the orthologs of the previously described mammalian
Eif4e1b gene (Evsikov et al., 2006) in the genomes of representatives for all major classes of
Tetrapoda. In all examined species, the origin of Eif4e1b gene is likely monophyletic, and its
expression is commonly upregulated in oocytes and ovaries, suggesting conserved function(s)
in female germline.
To further understand the evolutionary history of the Eif4e1b locus, we have performed the
TBLASTN search of all D. rerio ESTs and cDNA sequences to detect the eIF4E-coding
mRNAs in this species. Seven distinct eIF4E genes were identified, including previously
reported eif4e1a and eif4e1b (Robalino et al., 2004; Suppl. Table 1). Multiple sequence
alignment of corresponding proteins with eIF4E proteins of Tetrapoda using CLUSTALW
confirmed that four D. rerio genes, eif4e1a, eif4e1b, zgc:101581 (Eif4e1_2) and zgc:110154
(Eif4e1_3) encode proteins of the eIF4E1 subfamily (Suppl. Fig. 1). To our surprise, while the
zebrafish eIF4E1B is very similar to other eIF4E1B proteins and has a similarly restricted
pattern of expression (Robalino et al., 2004), the D. rerio eif4e1b gene is not orthologous to
either Eif4e1b or Eif4e loci of Tetrapoda. Indeed, alignment of eif4e1b cDNAs to the D.
rerio genome identified the eif4e1b locus unambiguously (Fig. 1C). However, although the
colinearity of flanking Slc7a2 and Casr genes is well-conserved, the genomes of other
Actinopterygii species (Gasterosteus aculeatus, Oryzias latipes, Takifugu rubripes and
Tetraodon nigroviridis) lack any sign of eIF4E-encoding locus in the respective regions (Fig.
1C). Conversely, while the physical linkages of Sncb and Tspan17 loci persist among these
four fish species and Tetrapoda, there is no Eif4e-like gene residing in this region of any
Actinopterygii genome as well (Fig. 1D vs. 1B). Together, these data indicate the lack of
orthology between the D. rerio eif4e1b and the Eif4e1b of Tetrapoda.
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Further TBLASTN search of fish genome assemblies using D. rerio eIF4E1A, eIF4E1B,
eIF4E1_2 and eIF4E1_3 protein sequences revealed three ostensibly orthologous groups of
eIF4E1-encoding genes in Actinopterygii (Fig. 2; Suppl. Table 1). The physical linkage of
Eif4e1_2 and Metap1 loci in all five species indicated Eif4e1_2 genes as 'genuine' orthologs
of Tetrapoda Eif4e genes (Fig. 2B vs. Fig. 1A), while Eif4e1_1 (Fig. 2A) and Eif4e1_3 (Fig.
2C) loci are unique to Actinopterygii species. To test these conclusions, multiple sequence
alignments of newly-identified eIF4E1_1, eIF4E1_2 and eIF4E1_3 proteins of G. aculeatus,
O. latipes, T. rubripes and T. nigroviridis were performed to identify their relationships to the
eIF4E1 family members of D. rerio and Tetrapoda (Suppl. Fig. 2). The results indicate that
the eIF4E1_2 proteins of Actinopterygii are indeed the closest relatives of Tetrapoda's eIF4E,
while eIF4E1_1 and eIF4E1_3 are distinct clades of this subfamily. Furthermore, eif4e1a gene
of D. rerio, absent in the (current) genome assembly, likely belongs to the Eif4e1_1 group,
while no close relative for eif4e1b gene could be identified in any other currently available
Actinopterygii genome. This may be due, in part, to the taxonomy of Actinopterygii, as D.
rerio has a very distant relationship to any of the other surveyed species (Suppl. Table 2).
Indeed, several likely eIF4E1B-encoding sequences were identified previously among cDNA
and EST sequences of other fish species (Joshi et al., 2005). However, absence of the genome
assemblies or even any considerable genomic sequence data precludes the studies of
orthological relationships for these additional eif4e1b genes.
Overall, our data strongly indicate rapid diversification of eIF4E1 subfamily occurred during
the time of whole-genome duplication in vertebrates (Nakatani et al., 2007). This diversity was
lost during the early phylogenesis of Tetrapoda lineage and resulted in the fixation of merely
two loci, the ubiquitous Eif4e and oocyte-restricted Eif4e1b, in these taxa. It is plausible that
multiple Eif4e1b-like loci existed in the common ancestor of Actinopterygii and Tetrapoda;
some of them were asymmetrically retained in Actinopterygii (e.g., eif4e1b locus in D. rerio),
while others, such as the ancestral locus that spawned Eif4e1b of Tetrapoda, were subsequently
lost in other lineages. Whatever the case, this provides an excellent example of convergent
molecular evolution for eIF4E1B-encoding genes which, while being phylogenetically distinct
in Actinopterygii vs. Tetrapoda, endure selective pressure to encode functionally identical
proteins, as well as to be preferentially expressed in female germline. In fact, this situation
strongly resembles the 'phylogenetic behavior' of some oocyte-expressed genes in mammals
that we discovered previously. For example, some critical maternal effect genes sustain
asymmetrical reduplications, and presumably strong selective pressure, to result in the
formation of multiple homologous, but not orthologous, genes belonging to several distinct
families (e.g., Nalp, Oasl, Oog and others; Evsikov et al., 2006).
To address the function of eIF4E1B protein in the oocytes, morpholino antisense
oligonucleotides (morpholinos) were injected to curb translation of its mRNA in the eggs of
X. tropicalis. Unlike Xenopus laevis whose allotetraploid genome allows each protein to be
translated from two different mRNAs and thus renders translational suppression experiments
unfeasible, X. tropicalis was the most suitable model to perform such studies because it has a
diploid genome and retains much of the advantages found in X. laevis as a model embryological
system (Carruthers and Stemple 2006). Although X. tropicalis stage VI oocytes express all
eIF4E-encoding genes identified in the genome of this species (Fig. 3A), we were interested
in the specific functions for the oocyte-specific isoform. Injection of anti-Eif4e1b morpholinos
into freshly harvested X. tropicalis fully-grown eggs per se did not result in any changes to
oocyte morphology for up to 144 hours of culture. This result implies that eIF4E1B protein is
dispensable at this stage and may function later in oogenesis, such as meiotic maturation, or
during early development. To test this hypothesis, oocytes injected with anti-Eif4e1b
morpholino oligonucleotides were induced to complete meiosis with progesterone and the rates
of meiotic maturation, measured by the appearance of 'white spots' on animal poles, the gross
morphological sign of metaphase spindle formation, were recorded. Since morpholinos only
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inhibit translation of cognate mRNA, progesterone treatment was applied to fully-grown
oocytes at 48 and 96 hours post-injection to ensure sufficient time for intrinsic protein turnover
of any pre-existing eIF4E1B proteins. Two days after injection, anti-Eif4e1b morpholino
treatment increased the number of matured oocytes at 3.5 hrs after progesterone treatment
(time×treatment χ2=49.6, P<0.0001) whereas no other time point was effected (Fig. 3B). Four
days after treatment, 2-to-8-fold more oocytes receiving anti-Eif4e1b morpholino treatment
(χ2=62.24, P<0.0001) completed maturation 30-60 min faster compared to all three control
groups (time×treatment χ2=58.96, P<0.0001), respectively (Fig. 3C). A similar effect of
meiotic acceleration by 30-60 minutes was obtained with oocytes induced with progesterone
at 144 hours post-injection (data not shown); however, in this case maturation in all groups
was delayed up to ∼7 hours, most likely due to the adverse effects of prolonged culture.
Interestingly, comparable acceleration of maturation was recently reported when the oocytes
from another frog, X. laevis, were injected with an antibody that recognizes a likely ortholog
of eIF4E1B in these species (Minshall et al., 2007). Overall, our data further underscores a
novel, unexpected conclusion that the oocyte-specific eIF4E1B acts as a translational
repressor of dormant maternal mRNAs. At first glance, this repressor activity is surprising, as
all eIF4E1B and eIF4E proteins from Tetrapoda and D. rerio are identical at 98 out of 183
residues constituting the IF4E/CDC33 domain (Suppl. Fig. 3). Moreover, all the residues
critical for 5′-cap mRNA binding and interactions with eIF4Gs or eIF4E-BPs are absolutely
conserved among eIF4E1Bs and eIF4Es (Suppl. Fig. 3). Despite this striking sequence identity,
our experimental data and the results from other laboratories suggest that functionally,
eIF4E1B is very different from 'canonical' eIF4E. Indeed, assuming the functional identity
among eIF4E1B proteins in vertebrates, the important characteristics of these proteins are weak
mRNA cap and eIF4G-binding, and strong affinity to 4E-T (Minshall et al., 2007; Robalino et
al., 2004), an eIF4E transporter protein encoded by Eif4enif1 gene in mammals. Recently, a
model based on the biochemical interactions of X. laevis eIF4E1B with 4E-T and CPEB in
immature oocytes has been proposed (Standart and Minshall, 2008). In this model eIF4E1B,
unlike 'canonical' eIF4E, behaves as translational co-repressor of CPE-containing maternal
transcripts required for maturation, such as Cyclin B mRNA. Importantly, this proposed model
may occur in mammalian systems as well because all three proteins are highly abundant in
mouse oocytes (Evsikov et al., 2006; Villaescusa et al., 2006). In the future, studies of the
biochemical interactions of eIF4E1B, which are beginning to be uncovered (Minshall et al.,
2007; Robalino et al., 2004; Standart and Minshall, 2008), will hopefully explain the exact
mechanisms and roles in translational control of maternal mRNAs.
Materials and Methods
National Center for Biotechnology Information (NCBI; Wheeler et al., 2008) BLAST engine
was used for all GenBank searches. ESTs and cDNAs were downloaded from NCBI. EST
sequences were assembled in consensus sequences as described (Evsikov et al., 2006), and the
encoded protein sequences identified using ORF finder (NCBI). Searches against the genome
assemblies for all species were performed using online search tools of ENSEMBL project
(Flicek et al., 2008; http://www.ensembl.org/). The following genome assemblies were used:
Anolis carolinensis, AnoCar1.0; Danio rerio, Zv7; Gallus gallus, WASHUC2; Gasterosteus
aculeatus, BROAD S1; Monodelphis domestica, monDom5; Mus musculus, NCBI m37;
Oryzias latipes, HdrR; Takifugu rubripes, FUGU 4.0; Tetraodon nigroviridis, TETRAODON
8.0; and Xenopus tropicalis, JGI 4.1. Multiple sequence alignments and their analyses were
performed using MEGA 4.0 (Tamura et al., 2007). The protein sequences of eIF4E family
members mentioned or identified in this study are provided in the Supplemental Table 3.
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Antisense Oligonucleotides
To inhibit translation of Eif4e1b mRNA, the 25-mer morpholino antisense oligonucleotides,
'morpholinos' (Gene Tools LLC, Philomath, OR, USA) were used. The sequence of anti-
Eif4e1b morpholino (5′CAGCTGCTGCCATTCTGCTATACAA-3′) was designed according
to manufacturer's recommendations using the 5′-end sequence of the X. tropicalis Eif4e1b
mRNA containing the AUG start codon (EST AL890091 and the corresponding region of X.
tropicalis genome assembly). This sequence does not match any mRNA but Xt_Eif4e1b, or
any other region of the sequenced genome. The 'standard control morpholino' (5′-
CCTCTTACCTCAGTTACAATTTATA-3′) is designed towards a splice-generating mutation
at position 705 β-globin pre-mRNA found in human thallasemia. This morpholino is commonly
used as a control in egg microinjection experiments and does not have any known target or any
significant biological activity in either X. laevis or X. tropicalis. Stock concentrations for both
morpholinos were 1mM in sterile water. For injection, 10 nl of 200 μM morpholino, or sterile
water, was injected per oocyte.
Oocyte collection, microinjection and maturation assays
Follicle-cell-free X. tropicalis stage VI oocytes were obtained by collagenase treatment (2%
type I collagenase, 0.1% SBTI, 0.1% BSA in 1× Modified Ringer's [MR] medium: 100 mM
NaCl, 1.8 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 5 mM Na-HEPES; pH adjusted to 7.8 with
NaOH) for 1 hour at room temperature with gentle shaking (De Sousa and Masui, 1990).
Oocytes were washed 10 times in 100 mM potassium phosphate buffer (28 mM K2HPO4, 72
mM KH2PO4, pH 6.5), 5 times in 1× MR, and sorted on a 100 mm agarose-coated Petri dish
in 1× MR. Oocytes were placed in the OR3 medium (50% Leibovitz's L-15 medium, 15 mM
HEPES, 100 μg/ml gentamycin, 50 U/ml mystatin, 50 U/ml penicillin, 50 μg/ml streptomycin)
for 2-3 hours at 25°C and processed for microinjection (Gallo et al., 1995). Batches of 25
oocytes were placed in 35 mm plastic Petri dishes with a piece of nylon mesh (0.8-1 mm
openings) attached to the bottom. Before injection, level of OR3 medium was adjusted to
200-400 μm above oocytes, so they were held in place by the mesh on the bottom and surface
tension on top. Injections were performed with a Picospritzer apparatus (Parker Hannifin Co.,
Cleveland, OH, USA) using needles pulled from glass borosilicate capillaries (outer diameters
of needle tips were 17-19 μm). After injection of morpholino or water (or no injection control),
the oocytes were cultured for 48, 96 or 144 hours in OR3 in agarose-coated Petri dishes at 25°
C. Maturation was induced in oocytes by incubation in 1 μg/ml progesterone in OR3 for 6–10
hours at 25°C. The meiotic progression was monitored by counting the number of oocytes with
the clearly defined white spot on the animal pole every 30 minutes beginning at 3 hours after
the start of progesterone treatment. Fifty oocytes per experimental group (anti-Eif4e1b
morpholino-injected, control morpholino-injected, water-injected and uninjected) in two
independent experiments conducted on different days were used.
Gene Expression
RNA was isolated from X. tropicalis fully-grown oocytes (a generous gift from Dr. Laurinda
Jaffe, University of Connecticut Health Center, Farmington, CT, USA). RNA was treated with
RNase-free DNase (Quiagen) and used for cDNA first strand synthesis (Superscript III,
Invitrogen). Conditions for PCR were optimized using FailSafe PCR system (Roche). The PCR
amplification program was: 97°C 1 min; 35 cycles of 95°C/30 sec; 58°C/30 sec; 72°C/45 sec;
72°C 10 min; 4°C infinity. PCR products were separated on 1% agarose gels, stained with
ethidium bromide and visualized by UV transillumination. Sequences of primers used for RT-
PCR studies are listed in Suppl. Table 4.
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Descriptive statistics (mean and SEM) were reported as the percent of mature oocytes at each
time interval. To detect statistical differences for the effects of treatment, time, and the
interaction of time × treatment on the total number of matured oocytes, an ordinal logistic fit
model was used to calculate the χ2 likelihood ratio tests, with differences considered significant
at P<0.05 (JMP 7 software, SAS Institute, Cary, NC).
Supplementary Material
Refer to Web version on PubMed Central for supplementary material.
We would like to thank Dr. Laurinda Jaffe for advice and for providing the opportunity to conduct the X. tropicalis
morpholino experiments in her laboratory at The University of Connecticut Health Center. This work was supported,
in part, by a grants from NIDDK (DK73267) and NIAAA (AA017244).
Bachvarova RF. A maternal tail of poly(A): the long and the short of it. Cell 1992;69:895–897. [PubMed:
Carruthers S, Stemple DL. Genetic and genomic prospects for Xenopus tropicalis research. Semin Cell
Dev Biol 2006;17:146–153. [PubMed: 16427790]
De Sousa PA, Masui Y. The effect of cytochalasin D on protein synthesis in Xenopus laevis oocytes. Mol
Reprod Dev 1990;26:248–252. [PubMed: 2375877]
Evsikov AV, Graber JH, Brockman JM, Hampl A, Holbrook AE, Singh P, Eppig JJ, Solter D, Knowles
BB. Cracking the egg: molecular dynamics and evolutionary aspects of the transition from the fully
grown oocyte to embryo. Genes Dev 2006;20:2713–2727. [PubMed: 17015433]
Flicek P, et al. Ensembl 2008. Nucleic Acids Res 2008;36:D707–D714. [PubMed: 18000006]
Gallo CJ, Hand AR, Jones TL, Jaffe LA. Stimulation of Xenopus oocyte maturation by inhibition of the
G-protein alpha S subunit, a component of the plasma membrane and yolk platelet membranes. J Cell
Biol 1995;130:275–284. [PubMed: 7615631]
Joshi B, Lee K, Maeder DL, Jagus R. Phylogenetic analysis of eIF4E-family members. BMC Evol Biol
2005;5:48. [PubMed: 16191198]
Minshall N, Reiter MH, Weil D, Standart N. CPEB interacts with an ovary-specific eIF4E and 4E-T in
early Xenopus oocytes. J Biol Chem 2007;282:37389–37401. [PubMed: 17942399]
Nakatani Y, Takeda H, Kohara Y, Morishita S. Reconstruction of the vertebrate ancestral genome reveals
dynamic genome reorganization in early vertebrates. Genome Res 2007;17:1254–1265. [PubMed:
Richter JD, Sonenberg N. Regulation of cap-dependent translation by eIF4E inhibitory proteins. Nature
2005;433:477–480. [PubMed: 15690031]
Robalino J, Joshi B, Fahrenkrug SC, Jagus R. Two zebrafish eIF4E family members are differentially
expressed and functionally divergent. J Biol Chem 2004;279:10532–10541. [PubMed: 14701818]
Standart N, Minshall N. Translational control in early development: CPEB, P-bodies and germinal
granules. Biochem Soc Trans 2008;36:671–676. [PubMed: 18631138]
Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA)
software version 4.0. Mol Biol Evol 2007;24:1596–1599. [PubMed: 17488738]
Villaescusa JC, Allard P, Carminati E, Kontogiannea M, Talarico D, Blasi F, Farookhi R, Verrotti AC.
Clast4, the murine homologue of human eIF4E-Transporter, is highly expressed in developing
oocytes and post-translationally modified at meiotic maturation. Gene 2006;367:101–109. [PubMed:
Wheeler DL, et al. Database resources of the National Center for Biotechnology Information. Nucleic
Acids Res 2008;36:D13–D21. [PubMed: 18045790]
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Figure 1. Conservation of genomic structure for Eif4e and Eif4e1b loci in Tetrapoda
Colinearity of genes underscores the common phylogenetic origins for Eif4e (a) and Eif4e1b
(b) in tetrapods. (c) The Danio rerio eif4e1b locus is not conserved among Actinopterygii
genomes. (d) While the colinearity of Sncb and Tspan17 orthologs persists across vertebrates,
fish genomes lack the Eif4e1b locus. Orthologs shown in the same color; non-orthologous
genes shown in black.
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Figure 2. Actinopterigyii genomes contain multiple members of eIF4E1 subfamily
Danio rerio expresses at least four members of this family (Suppl. Table 1), three of which
have orthologs in other sequenced fish genomes. Genomic arrangement of loci for putative
orthologs of zebrafish eif4e1a (Eif4e1_1) (a), zgc:101581 (Eif4e1_2) (b) and zgc:110154
(Eif4e1_3) (c) genes. Orthologs shown in the same color; non-orthologous genes shown in
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Figure 3. Gene expression of Eif4e1b and its function in oocytes of Xenopus tropicalis
(a) RT-PCR detection of eIF4E-encoding genes (Xt_Eif4e1b, Xt_Eif4e, Xt_Eif4e2a,
Xt_Eif4e2b, Xt_Eif4e3), Xt-Gpcr13, Xt-Odc as positive controls of cDNA quality and H2O as
a negative control. (b, c) Effects of blocking eIF4E1B translation on the resumption of meiosis
and maturation in oocytes exposed to anti-Eif4e1b morpholino or control for two and four days.
Evsikov and Marín de EvsikovaPage 10
Dev Genes Evol. Author manuscript; available in PMC 2010 May 4.
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text:
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Fish Market in Ellsworth ME or surrounding area?
- whs Jul 6, 2008 06:35 PM
We're spending a long weekend with friends who have rented a house in Sullivan ME. We are charged with cooking dinner for a 15-20 people and thinking of doing grilled bluefish. Any recs for a good fish market in the area?
Dunno about bluefish but try Grindstone Neck in Winter Harbor - www. grindstoneneck.com or 1-888-831-8734. Carl Johnson, who runs a fish smoke house and a restaurant in Winter Harbor is selling fresh seafood from his smoke house in Winter Harbor. Last I saw he had great halibut, coho salmon, flounder, etc. Give him a call. He might be able to get in some bluefish or, in fact, suggest some other great stuff.
re: Tipatina
We know Carl--thanks for the suggestion.
Pectic Seafood, long established in Halls Quarry on MDI, just opened a huge store with cafe on Rte. 3 in Trenton: http://www.pecticseaffood.com
There's also a wholesale place in Ellsworth that might be of some help. Maine Shellfish. 207-667-5336 or 800-666-5336.
Got to check out Pectic Seafood, Mainegal. Thanks.
re: Passadumkeg
Actually, our friend who lives in West Gouldsboro suggested going to Hannaford in Ellsworth--is their fish any good?
re: BikeToEat
Ells. Hannaford, has good fish, but not fresh off the boat fresh. I don't know if this is still possible, one used to be able a couple of years a go to meet the 2 Bar Harbor ground fishing boats ant the town landing on Fridays (They go out on Mondays.) and buy fish fresh off the boat.
It's ok, not thrilling, but... Always lots of Farm-Raised Salmon, sometimes Artic Char, always haddock either fresh or FAS, that kind of thing. Sometimes they do get great fresh swordfish. You might want to call and see what they could do for you - special order or whatever for that many people. 207-667-5300. Do think you'd get better quality through Carl Johnson who is always eager to be helpful. Might have some good ideas for feeding your hoardes.
re: Tipatina
I sent him an email but have yet to hear back.
re: Tipatina
Friday. Tell him it's the guy who grew up in Thailand that liked the painting of the school kids in NE Thailand that hung in the hallway of the B&B.
re: Tipatina
Yikes, close. Try calling -- 207-963-7347. He (Carl) seems to be there, at the smokehouse, in the morning, but has great people even when he's not there who will surely take a message.
re: Tipatina
thanks, we have a camp off 182 on Tunk Lk. and visit Tibbet's often.
re: Passadumkeg
She said that the overhead in Milbridge was too much, so went back to the original place. When I was there, they were just getting started but looks like she'll be having pretty much the same inventory. Haddock, salmon, crab meat, lobster, crab legs, etc. Mr. Tibbet's had just arrived with clams! It's before you get to Country Charm so don't despair if you think you've gone too far.
re: Tipatina
Do you come up for summers or live year round? What town(s)?
re: Passadumkeg
Now May to October. Gouldsboro near the Winter Harbor line. Moved from NY to Yarmouth to be closer. Love it so much. You're Bar Harbor in the winter?
BTW, August 11th, Bill Thayer from Darthia Farm will be playing with his jazz group at Tidal Falls.
re: Tipatina
Thanks, will try to make it. No I live year round and teach in Ellsworth, work as a sea kayak guide around BH for the summer, used to live in SWH and N Haven Island, spent long time overseas.
To the Topic. There is a fresh fish wagon on High St in Ellsworth, next to Irvings.
Hve you tried Chester Pike's Galley fpr break.? Lobster or Chester Benedict, best corned beef hash I know.
Thanks everyone for the suggestions--we ended up at Pectic Seafood, which had a small but very fresh selection of fish. No bluefish, we ended up making Jasper White's Portuguese-style chowder with chourico, haddock and hake. BTW, they had garlic scapes, which we added to the stew. They also carry a number of refrigerated items--we found Duck Trap salmon ends for $10 a pound--a bargain!


text:
> Home
> Calendar
> Weekday Hike - Theodore Roosevelt Area
Feb 14, 2012
10:00 a.m.
Feb 14, 2012
10:00 a.m.
Explore Timucuan Preserve along the St. Johns on hiking trails through hardwood forest, marshes, bluffs, and shell mounds. No pets.
Contact Information:
Northeast Florida Group
Bill Armstrong, email@example.com
Northeast Florida Group
Bill Armstrong, firstname.lastname@example.com
NEAR: Jacksonville
The trail entrance is at 13165 Mount Pleasant Rd., 1 mile southeast of Fort Caroline National Memorial. Bring water and insect repellent. Children over 8 OK. Coming from the west of Fort Caroline National Memorial on Mount Pleasant Rd., pass the entrance, go about 2 miles more on Mount Pleasant Rd, and turn left at the brown sign on a cyclone fence on the left that says "Trailhead Parking". Coming from the east of the Memorial, pass signs for Spanish Pond development and go 1/4-1/2 mile on the right. Brown signs are flat on the cyclone fence.
NEAR: Jacksonville
Read more at this website
Copyright Sierra Club Northeast Florida Group


text:
Nippon Yusen K.K.'s New Car Fleet Tour
Similar imagesView all
A worker walks near the Nippon Yusen KK's car transporter ship Auriga... News Photo 116257651Asia,Asian,Business,Car,City,Corporate Business,Factory,Finance,Horizontal,Industry,Japan,Japanese,Kanagawa Prefecture,Mitsubishi Heavy Industries,Mitsubishi Motors,Mode of Transport,Occupation,Ship,Shipping,Walking,YokohamaPhotographer BloombergCollection: Bloomberg 2011 BloombergA worker walks near the Nippon Yusen K.K.'s car transporter ship, Auriga Leader, at Mitsubishi Heavy Industries Ltd.'s Honmoku plant in Yokohama city, Kanagawa prefecture, Japan, on Wednesday, June 15, 2011. Nippon Yusen K.K. is Japan's largest shipping line. Photographer: Tomohiro Ohsumi/Bloomberg via Getty Images


text:
AJOT Digital Edition
Issue #592
Breakbulk Quarterly
3PL Quarterly
|2014 Media Kit
Transport Intermediaries
Page 73 of 130 pages ‹ First < 71 72 73 74 75 > Last ›
Damco extends Cargo 2000's coverage of the global air cargo market
| April 03, 2012 | Transport Intermediaries
Already representing over 70% of the global air cargo market, Cargo 2000's quality management system has been strengthened further by Damco's decision to join the group. Read More »
FPS Hawaii groupage cuts time and cost
| April 03, 2012 | Transport Intermediaries
FPS Hong Kong, one of the founder members of the Famous Pacific Shipping Group forwarding and logistics network , is launching a new direct ocean groupage service to Honolulu, Hawaii that reduces traditional costs and transit times. Read More »
Agility Q4 net profit rises 114%
| April 01, 2012 | Transport Intermediaries
Kuwait's Agility, the logistics firm facing U.S. fraud charges, posted a 114-percent rise in fourth-quarter net profit compared with the same period in 2010, the firm said in a statement. Read More »
Data2Logistics certified as TRUSTe EU Safe Harbor amid global growth
| April 01, 2012 | Transport Intermediaries
Global freight bill processor, auditor and advisor Data2Logistics announces the designation of the organization as a TRUSTe EU Safe Harbor Certified Company. The program verifies a company's compliance with the EU Safe Harbor Framework, and verifies self- Read More »
Globe Express Services (Overseas Group) launches joint venture with ITSA (Italy)
| April 01, 2012 | Transport Intermediaries
Globe Express Services' (Overseas Group), one of the world's top 100 global logistics providers, recently finalized an historic joint venture (JV) with International Transport Services Associated (ITSA). Headquartered in Rome, Italy, ITSA has spent over 3 Read More »
RedStone Logistics offers new StoneBench service, a diagnostic transportation assessment tool
| April 01, 2012 | Transport Intermediaries
RedStone Logistics, a third-party logistics company specializing in logistics management services, has begun offering a new diagnostic transportation assessment service, StoneBench. The new service, provided at no cost for a limited time, provides compani Read More »
Shipco adds operations office in Laem Chabang, Thailand
| April 01, 2012 | Transport Intermediaries
Shipco Transport has announced the opening of its third operations office in Thailand. Situated in Thailand's largest port, Laem Chabang is located 120 km southeast of downtown Bangkok and 90 km southeast of Lat Krabang and Suvarnabhumi airport. The new o Read More »
Supply chain consulting firm expanding headquarters in Hamilton County
| March 29, 2012 | Transport Intermediaries
enVista, LLC, an enterprise and supply chain consulting services firm, announced plans to expand its headquarters here, creating up to 96 new jobs by 2016. The company, which serves customers in the retail, automotive and health care industr Read More »
U-Freight partner gains AEO status
| March 29, 2012 | Transport Intermediaries
Tracosa Aduanas, the Spanish independent freight forwarder and agent of the U-Freight Group (UFL), has become an Authorised Economic Operator (AEO) in the Customs Simplifications category. Read More »
Hellmann Worldwide Logistics first logistics service provider to join Sustainable Apparel Coalition
| March 28, 2012 | Transport Intermediaries
The Sustainable Apparel Coalition (SAC) is a coalition of leading apparel and footwear companies who represent over 30% of the industry's global market share. SAC members promote a common approach to measuring and evaluating the social- and environmental Read More »
DSV has been fined 379,000 euros by EU
| March 27, 2012 | Transport Intermediaries
Danish freight forwarder DSV said the European Commission had ordered it to pay a fine of 379,000 euros ($504,900) for a subsidiary's participation in a European cartel. Read More »
EU sets cartel fines on freight forwarding firms of 169 million euros
| March 27, 2012 | Transport Intermediaries
Thirteen logistics firms, including UPS, Panalpina and Expeditors, were fined a total 169 million euros ($225 million) by the European Commission for cartel activities. Read More »
Food aid graft worsened Kenya drought impact
| March 27, 2012 | Transport Intermediaries
Rampant corruption in food aid worsened the suffering of Kenyans hit by drought last year, Transparency International said, with some politicians sabotaging food distribution and giving it out where it was not needed to win support. Read More »
Innovative rolling loading dock safety gate offers OSHA compliant fall protection
| March 27, 2012 | Transport Intermediaries
Diversified Fall Protection recently unveiled an improved loading dock safety gate to further complement the company's comprehensive line of fall protection solutions. This OSHA-compliant rolling safety gate protects personnel from loading dock falls and Read More »
Kuehne & Nagel mull appeal, after EU sets cartel fine
| March 27, 2012 | Transport Intermediaries
Swiss logistics group Kuehne & Nagel said it is considering appealing against an 53.7 million euro ($71.54 mln) fine from the EU commission decision for cartel activities, saying the investigation was not carried out correctly. Read More »
EU sets cartel fines on freight forwarding firms of 169 million euros
| March 27, 2012 | Transport Intermediaries
Thirteen logistics firms, including UPS, Panalpina and Expeditors, were fined a total 169 million euros ($225 million) by the European Commission for cartel activities. Read More »
Menlo Worldwide Logistics appoints two new directors in South Asia
| March 26, 2012 | Transport Intermediaries
Menlo Worldwide Logistics, the global logistics subsidiary of Con-way Inc., has announced the appointments of two new directors within its South Asia organization. Victor Loo was named director of Operations, while Kelza Phang has been appointed director Read More »
PSS Distribution Services chosen as Juan Valdez's' 3PL transportation services provider
| March 26, 2012 | Transport Intermediaries
3PL warehousing and transportation company PSS Distribution Services has agreed in principal to provide distribution services for Juan Valdez', a company whose name is synonymous with Colombian coffee and is well established in the coffee industry. Read More »
EU to fine UPS, other logistics firms over cartel
| March 26, 2012 | Transport Intermediaries
EU antitrust regulators are set to fine 13 logistics firms, including UPS, Panalpina and Expeditors, for taking part in a cartel, two people familiar with the matter said. Read More »
Technology, equipment benefits perishables imports
| March 22, 2012 | Intermodal | Transport Intermediaries
Supply chain logistics for perishables is a growth industry facing many challenges. For starters, last November the world's population hit over 7 billion. Half of that population was added in only a short 40 years, and 3 billion more people are expected to inhabit the earth by 2100, bringing population figures to 10 billion. Read More »


text:
Based on 1 review
Help make it easier for other Avvo users to choose the right lawyer by sharing your experience with this attorney. It's fast, simple, and safe.
Lawyers: Use the Peer Endorsements section to provide input about other attorneys.
I've dealt with Laurel exclusively for over 2 years on my increasingly complex child custody and divorce case. She has been superbly knowledgeable about every aspect of my case including mediation, courtroom litigation, dealing with expert witnesses, medical issues, multiple levels of appeals including the NC Supreme Court, cross state jurisdictional situations, equitable distribution, alimony, child support.....you name it, Laurel can handle it with utmost professionalism.
In addition to her superior legal knowledge Laurel is kind, sympathetic, someone who truly cares not only for you and your case but also for what is in the best interests of, in my case, the children involved. She actively seeks to involve you in decisions about your case so that you are an active participant and a true partner in resolving your case.
I truly cannot say enough positive things about Laurel's legal capabilities and personal integrity. If you need legal counsel you will do no better.


text:
1951 Chevy 4 Dr Sedan, 3 speed manual transmission, 6cyl in-line 37,473 miles... More
1958 Mercedes Benz 190 SL Convertible Details available soon ... More
1959 OLDSMOBILE DYNAMIC 88; RESTORED TO VERY HIGH STANDARDS, THIS ONE OF A KIND OLDSMOBILE. POWERED BY A 420 H.P. CADILLAC 6.0 ENGINE WITH... More
1953 MG TD http://www.ftlauderdaleauction.com/... More
Olde Tyme Auctions - Outstanding Sale 1957 Chevy Bel Air - and other items! Saturday; November 29, 2014 4:00 pm Doors open at 9:00 am for Preview 41... More
1951 Ford Woody Complete frame off no expense spared restoration. All new wood & Mahogany veneer from the Honduras. Painted 2000 Jaguar Carnival Red with... More
1956 Packard Caribbean Convertible The Caribbean by Packard was one of the signature cars of the 1950's. Only 276 Caribbean convertibles were built. They were... More
1952 Hudson Hornet Twin H 160HP Stick over drive. This model featured the step down chassis design which provided greater passenger safety as well as... More
- New steering box- Springs and shackles- New Coker whitewall tiresTo be sold at the Kansas City 2014 auction, Kansas City, MO, December 4-6,... More
- Viper Red with Tan leather and cloth interior- Restored in 2013 - 149 miles since restoration- New 350/420 HP roller engine- Ford rear... More
- Body off restoration- Fuel injectedTo be sold at the Austin 2014 auction, Austin, TX, December 12-13, 2014. To consign a car or to... More
- Electric windows- Continental Kit- Rocker moldings- 2-door convertible- Bumper wings- Wire wheelsTo be sold at the Austin 2014 auction, Austin, TX, December 12-13,... More
To be sold at the Kissimmee 2015 auction, Kissimmee, FL, January 16-25, 2015. To consign a car or to register to bid, visit the... More
- 3-speed transmission- Same owner since 1995- Rare Highland Green paint- Green interiorTo be sold at the Kissimmee 2015 auction, Kissimmee, FL, January 16-25,... More
- 350 CI crate engine- 5 window California truck- Turbo 350 automatic transmission- Fatman Fabrication Mustang II front end- Late model rear- Power brakes-... More


text:
The Top Six New Diets
While veganism is restrictive, "the best way to approach it isn't by cutting things out," says Jack Norris, R.D., author and founder of Vegan Outreach, "it's by adding them in." First, make sure you include ample plant foods at each meal, especially ones high in protein such as soy meat, tofu, beans, falafel, and nuts. The idea is that if you load up on vegan foods, you'll naturally eat fewer animal products. In spite of his own commitment to veganism, Norris doesn't advocate rigidity. "I tell people to avoid obviously animal-based items, but don't quibble over every ingredient. You don't need to quiz the wait staff about whether there's egg in the pasta or traces of dairy in the dinner rolls."
As for ensuring adequate protein intake on a vegan diet, Norris says that if you eat a variety of vegetables, legumes, fruits, and grains, you'll likely get enough. "It's a myth that you need to get all your amino acids at one sitting. Your body collects different ones from different foods and assimilates them as needed," he says.
If veganism seems too strict but you like the idea of eliminating most animal products, consider one of the following alternatives:
Lacto vegetarian: eats dairy products
Ovo vegetarian: eats eggs
Lacto-ovo vegetarian: eats dairy and eggs
Pollotarian: eats poultry and fowl
Pescatarian: eats fish and seafood
To learn more about how to transition to a vegan diet, visit jacknorrisrd.com.
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Add YOUR event today! It's easy to do, and completely FREE! ADD YOUR EVENT
Been coming here for years. They consistently have delicious dim sum, though some items are not as great quality (chicken feet). I always love the baozi...Read More
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text:
didnt say it was a bad product. there is more to a product than the item itself. i know what i get with the bcm stuff which stupid fast after the sale service if something does go wrong. i dont know what or how fast spikes does cs turn around.
on paper they look very similar.
so you got to dig a bit deeper for differences. do you have experience with spikes cs??
another difference is spike's has an ugly logo printed on the carrier :P plus they are in florida not wisconsin. something suspicous about that...
"Gun control: The theory that a woman found dead in an alley, raped and strangled with her panty hose, is somehow morally superior to a woman explaining to police how her attacker got that fatal bullet wound." L. Neil Smith
The problem with being stupid is you cannot simply decide to stop doing dumb things...
Sent from my iPhone using Firearms Talk


text:
Senior talent on display
From behind a sparkling silver curtain June 30 emerged Eileen Grant in a red jacket and black fedora, her left hand gloved in silver, dancing and swinging her hips to Michael Jackson's "Bad," while fellow residents of the Emeritus at Deep Run retirement home and community members laughed and clapped.
Grant and other residents, volunteers and community members were participating in the first national Bring Your Talent contest, hosted by Emeritus Senior Living and part of elder care expert Dr. Marion Somers' tour with the 3in4 Association, a non-profit organization working to increase awareness about the planning for long-term care. The 50 city tour will end July 25 in Phoenix, A.Z. and the contest winner will be chosen from all the video submissions and taped tour stops by Aug. 13. The winner could receive a year of free rent or one of 11 week-long stays at an Emeritus community.
Grant was born in 1928 in Brooklyn, N.Y. as Eileen O'Connor, the third-oldest of 10 children in an Irish family. During World War II she traveled with other dancers and comedians and pros such as Mel Brooks and Eddie Fisher entertaining the troops.
"I loved it. I was doing it from the time I was this high," Grant said, lowering her hand toward the floor. "If you're a performer, you're born a performer."
Grant's daughter, Shelley Damion, who attended the show last week but sat on the side so as to not to be a distraction, said she hoped her mother would win.
"She's always been the most entertaining, even at parties," Damion said. "She choreographed my high school production of 'Bye Bye Birdie.' I think it was the best one we had ... I wasn't in it."
Every Friday, Grant, who is a memory care patient, practices ballroom dancing with instructor Phyllis Harris, who helped co-choreograph her dance.
"It's really exciting to see them come out of their shells," Harris said. "It's hard to watch them get older. . . She comes down when she feels like it."
Harris was supposed to perform with Grant the day of the show, but because other entries dropped out, Grant had to perform before Harris had arrived. Filling in for Harris was Nick Nevi, the life enrichment assistant at Emeritus, and one of Grant's biggest fans.
"She's got spunk," he said the day before the contest. "She's very talented. She's my hopeful. I know she could really knock the house down. . . She deserves one year rent free."
Nevi said he hoped that if she didn't win, that one of the other Deep Run residents would.
"When our residents do what they love it, rejuvenates them," he said. "I really admire our residents for doing this."
The Thursday before the contest, Harris and Nevi made a video music video of Grant's performance in case she got nervous the day of or resisted because of her memory.
Private duty caregiver Demetra White said she had seen the end of the videotaping.
"When I saw her get up and these moves, she seemed so much younger, " White said. "My mouth was just open. If she doesn't win, I'll be surprised."
During the practice, Grant pulled White onto the dance floor despite White's protests. By then, Grant had kicked off her shoes and was dancing barefoot with free-style moves, something White said Grant usually did.
Old photos of Grant dancing and acting in the 1940s lined the walls of the dining room, where the stage was set for the performance on Saturday. In one or two, Grant's bare feet could be seen. Grant said she had learned to dance after attending dances with soldiers on Governors Island, where her older sister had a job.
"It was the kind of music where we'd go dancing and the gentleman would swing me over his head, around his back and between his legs," she said. "I was young and had a wonderful time. . . It's been a lot of years, but I had a wonderful life. I loved the life I lived. Isn't that a song?"
In her opening speech Saturday, Somers introduced herself as a girl from a rough neighborhood in East Harlem, N.Y., who grew up in a five-story walk-up filled with seniors. Somers said that there was always something to learn from her elders, especially how to live.
"You don't get to be 80, 90, or 100 years old without knowing how to live," she said.
On a trip to China, Somers met a 104-year-old woman who she asked to share with her one thing she'd learned. The woman responded, "Don't let the bastards get you down."
"I asked the interpreter what she meant," Somers said. "What she meant was, your husbands and kids are going to do what they're going to do, just do what you can do."
Having a balanced and steady approach to addressing long-term care is key, as it is to addressing life, she said.
"I like to say that I have been blessed living with seniors," Somers said. "I live a very blessed life because I've learned not to get obsessed about the nonsense stuff, and when you think about it, most of it is nonsense."
Somers went on to emphasize that everyone is a caregiver at some time in his or her life, whether it is for a little sister or for parents.
"Three in four Americans will need long-term care at sometime in their lives, and we are not ready," Somers said.
Sixteen percent of caregivers die before those for whom they are caring, she said, emphasizing the importance of being organized during the caregiving process. She went through an A-Z list of quick things for care givers to remember; among them: "Trust the elder you're caring for, and trust your basic caregiving instincts," and "Understand that being a caregiver requires patience."
"Talking about elderly care is not a sexy subject, so to remind people is necessary," she said. "Nobody wants to talk about aging or maybe being infirm. It's always over there, another person, but that other person could be you."
MADD to host candlelight vigil Dec. 2 at UR
Mothers Against Drunk Driving (MADD) will host a candlelight vigil of remembrance and hope Tuesday, Dec. 2 at 7 p.m. at the University of Richmond, outside the Cannon Chapel. The public is invited to attend and join MADD to honor victims of impaired driving crashes, while helping to remind the community to be safe during the holidays. > Read more.
Tournament supports adoption efforts
Among participants at the Seventh Annual Coordinators2Inc Golf Tournament and awards luncheon Oct. 3 were (from left) Rebecca Ricardo, C2 Inc executive director; Kevin Derr, member of the winning foursome; Sharon Richardson, C2 Inc founder; and Frank Ridgway and Jon King, members of the winning foursome.
Held at The Crossings Golf Club, the tournament will benefit placement of children from Virginia's foster care system into permanent families through Coordinators2. > Read more.
A.C. Moore to host winter craft day for kids
Event will help kick of Marine Corps' 'Toys for Tots' campaign
All 140 A.C. Moore locations will serve as drop-off centers this year for the Marine Toys for Tots Foundation, and all toys collected will stay in the local communities served by the stores in which they are donated.
On Saturday, Nov. 15, the Willow Lawn location will kick off the month-long program by hosting a "Make & Take" craft event for kids. Children ages six and older will be able to make a craft and take it home with them. Representatives from the Marines will be in-store to teach customers about the Toys for Tots program. A.C. Moore team members will be on site to help with the crafts. > Read more.
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Authentically Italian
Bella's feels – and tastes – like Italy should
Short Pump is known for its share of chain restaurants and strip malls, but diners looking for something more distinct can certainly find it without heading downtown or to nearby Charlottesville.
In fact, local husband-and-wife restaurateurs Valeria Bisenti and Doug Muir brought a taste of Charlottesville (and Italy) to Short Pump when they took a chance and opened Bella's second location in the same shopping strip as Wal-Mart and Peter Chang China Cafe. (Bella's original location is on Main Street in downtown Charlottesville.)
For a local Italian restaurant, Bella's is as "Mom and Pop" as its gets. Valeria is Mom, and Doug is Pop. Since its opening about six months ago, diners have been eating rich comfort foods and drinking Italian wines. > Read more.
Restaurant watch
Find out how your favorite dining establishments fared during their most recent inspections by the Virginia Department of Health. > Read more.
'Sizing Up!' opens at Cultural Arts Center
The Cultural Arts Center unveils a new exhibit – "Sizing Up!" – Nov. 20-Jan. 18 in the Gumenick Family Gallery.
Artist Chuck Larivey has spent the past three years "sizing up" – creating large-scale oil paintings that are designed to engage their viewers in a monumental way by using size to captivate them and make them a part of the artistic experience.
The exhibit is appropriate for all ages and is free and open to the public at the center, located at 2880 Mountain Road in Glen Allen. > Read more.
Stowe honored with Procurement Xcelerator Award from Virginia General Services
Willow Lawn to offer holiday "trolley" service from Church Hill
Student helps sprout 'STEM' growth
Grapevine access opens
McDonald's operator wins national award
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Nov. 20, 2014Click here to read the print edition.
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Weekend Top 10
A 'Life' well lived
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Obituary – John W. Wimble
Obituary – Sandra Stone Gillum
Obituary – Vivian Eleanor Muldoon
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Tournament supports adoption efforts
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Richmond Strikers form girls lacrosse program
Virginia fisherman wins James River event in Henrico
Hermitage to host Monday night football events
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Bette Midler: 'Ariana Grande looks ridiculous'
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Even more cuts at United Mileage Plus
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Kate Hudson introduces workout line
Actress Kate Hudson has stepped into the world of fashion by introducing a collection of workout clothes.
"The Something Borrowed" star has co-founded the gym collection, titled Fabletics, with online fashion retailer JustFab Inc.
The actress, who encourages women to exercise in style, hopes the line, which includes sportswear and accessories, will be affordable as all items are priced under $100.
Hudson tells the New York Daily News, "At Fabletics, we believe all women should be able to have hip styles and amazing quality at prices that won't break the bank."
The collection, which features sports bras, jackets, sweatshirts and scarves, is available online through the shopping website.
Kate Hudson taking secret dance lessons
Actress Kate Hudson has been brushing up on her fancy footwork by taking secret lessons with professional dancer Derek Hough.
The "Almost Famous" beauty showed off her skills during her recent stint as a dance teacher on hit TV musical Glee, but she has been using her free time this summer to perfect her moves with Dancing With the Stars regular Hough.
The choreographer tells E! News, "I've been working with Kate Hudson a little bit lately for fun – just dancing for fun, giving her some lessons. She's become a good friend and she's great."
However, Hough insists the 34-year-old has no intention of ever testing out her skills on the popular ballroom dancing competition.
He says, "I would love for her to do the show, but she would never. She's fantastic. But I want her to maybe do a guest spot."
Kate Hudson breastfed on the 'Reluctant Fundamentalist' set
Actress Kate Hudson is thankful director Mira Nair was so accommodating while filming "The Reluctant Fundamentalist" because she had to frequently take breaks to breastfeed her baby son.
The actress gave birth to Bingham, her first child with fiancé Matt Bellamy, just two months before she began shooting the thriller, and Hudson has heaped praise on filmmaker Nair for making time for her mum duties.
During an appearance on "Live! With Kelly and Michael" on Tuesday, she said, "I was hiding in cylinders on set breastfeeding Bing and it was a really intense role so I was going from breastfeeding to bawling my eyes out. But it was a really amazing experience.
"Mira Nair is an amazing director, she's very maternal and sensual and she creates this very passionate vibe on set. And so she's rubbing my feet, talking about the script, as I'm breastfeeding Bing, on set, so it was really interesting. It was a really wonderful working experience."
Glam night out at the Tiffany & Co Blue Book Ball
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Gwyneth Paltrow missed the "Iron Man 3″ premiere in London on Thursday night and was instead spotted partying in New York City at the Tiffany & Co Blue Book Ball. Other stars who turned out for the glamorous party in Manhattan included Kate Hudson, Carey Mulligan and Sarah Jessica Parker, while Robert Downey, Jr. headed the "Iron Man" cast without his leading lady in London.
Elizabeth Hurley and Kate Hudson support Elton John at breast cancer gala
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Elizabeth Hurley and Kate Hudson were among the glamorous guests at the annual Breast Cancer Research Foundation gala featuring Elton John in New York City on Wednesday.
The pair walked the fuschia-colored carpet for the Hot Pink Party at the Waldorf Astoria hotel in Manhattan, where British actress/model Hurley acted as the evening's emcee.
Hudson was on hand to present the Sandra Taub Humanitarian Award to Kay Krill, who heads retail company ANN INC., which is renowned for helping fund breast cancer research.
Sir Elton then thrilled revellers by performing some of his classic tracks at the charity bash, which was also attended by Hudson's Muse rocker boyfriend Matt Bellamy.
Kate Hudson was worried about Matt Bellamy's reaction to baby news
Actress Kate Hudson feared her surprise pregnancy would put a strain on her relationship with Muse rocker Matt Bellamy because she discovered the "shocking" news just two months into their romance.
The star announced she was expecting her first child with Bellamy in January, 2011 and the Hollywood beauty admits it was a stressful time for the new couple.
She tells Britain's Elle magazine, "It was all very old-fashioned and proper and we went on lovely dates, but two months later I was pregnant. I was shocked, but I also thought: 'This is going to be interesting.' I knew it would take a strong man to deal with it.
"You know that with a relationship things can go either way when you have a child. When you get pregnant everything changes; you, your body. Everything becomes a big decision, but he was there and that was that."
The pair became engaged three months later and that summer Hudson, who also has a 9-year-old son with ex-husband Chris Robinson, gave birth to a boy, Bingham – and the couple is still together.
She adds, "Now it's three wonderful years later. Things happen for a reason."
Hudson insists her relationship with Bellamy is stronger than ever, but they likely won't wed in the near future: "We will get married. I do think it's important but we have no plans. Ryder wants a party."
Kate Hudson's healthy diet down to Goldie Hawn
Kate Hudson has credited her mother Goldie Hawn with helping her maintain a healthy lifestyle, as the veteran actress advised her from a young age not to become "obsessed" with her weight or embark on extreme diets.
The "Almost Famous" star keeps her slim figure in check by avoiding wheat and dairy products, and reveals it was a chat with Hawn which helped her create realistic food habits which would benefit her as an adult.
She is quoted by Britain's Daily Express as saying, "When I was younger my mom advised me not to start going on crash diets or become obsessed with my weight and I have always stuck by that.
"I started eating properly at the age of 16, getting on a regime in which I'd get more energy. So I don't eat wheat or dairy. It is really simple. I became used to it very quickly and I don't crave stuff. I occasionally cheat, with a sandwich, which has great French bread and cheese, but not often. But mom is full of great advice – she's pretty handy like that."
Kate Hudson turned to mom for advice on juggling career, parenthood
Actress Kate Hudson turned to Goldie Hawn for advice on juggling her career with parenthood because the Oscar winner continued to find success even after becoming a mother.
Hudson hasn't slowed down since welcoming sons Ryder, 9, and Bingham, 19 months, and she is currently busy shooting a guest arc on "Glee."
Hudson admits she owes her strong work ethic to her mother Hawn, who showed her it's possible to balance life as an actress with motherhood.
She tells Glamour magazine, "I always remember my mom saying that there's only one thing in life she refused to fail at: raising her children. It just shows you it's very possible to do the thing that makes you happy and still raise a wonderful family."
Hudson insists she is enjoying every minute as a mother and reveals she has been able to keep her svelte physique by chasing after her youngest son, who loves to run around.
She gushes, "He's sprinting! And he's got a natural mullet, which I'm cutting soon. It's growing only in the back, so we in the family say he takes after… Kurt (Russell, my stepfather) because Kurt was a mullet guy in the eighties. Bing's been singing too. He loves music. And Ryder is the coolest cat – an amazing big brother, so patient and tolerant. Bing pulls his hair, and Ryder just laughs. He's such a good person. I couldn't be a prouder mommy."
Kate Hudson snaps and sings at Muse's London benefit
Actress Kate Hudson turned concert photographer on Monday night as she captured fiancé Matt Bellamy's band Muse live at their War Child Brit Awards concert in London.
The "Almost Famous" star joined Rita Ora, One Direction singer Harry Styles and Kate Moss' husband Jamie Hince at the O2 Shepherd's Bush Empire for the gig, and was spotted shooting the band from the stage and singing along to tunes like "Supermassive Black Hole" and the official 2012 Olympics anthem, "Survival."
The concert was Muse's smallest London show in a decade and won the band rave reviews.
Kate Hudson called Lea Michele to dismiss feud rumors
Actress Lea Michele was pleasantly surprised to receive a personal phone call from "Glee" guest star Kate Hudson recently after false tabloid reports suggested the pair had fallen out.
The "Almost Famous" beauty played sexy dance instructor Cassandra, a teacher at the New York school attended by Michele's character Rachel Berry, on season four of the hit musical comedy, but gossip from the set alleged Hudson was not a fan of the 26-year-old.
The unexpected rumors came as a surprise for both ladies, but Hudson made sure the young star knew how much she adored her by picking up the phone to dismiss the claims herself.
Recalling the phone call, Michele tells America's Marie Claire magazine, "(She said), 'I just heard about that (the rumored feud), and it [bleep] sucks. I want you to know I think you're amazing.'"
The touching conversation turned into a bonding moment for the co-stars and it was a call Michele will treasure for years to come.
She says, "It was so classy of her to take a minute to (debunk it), even though we both knew it was the biggest piece of crap in the whole world. I told her, 'I love you, and I adore you. Thank you so much for creating a human moment with me.'"
Michele admits tabloid gossip is "super-frustrating sometimes", but she has learned to just let it go when the magazines get it really wrong.
She adds, "Now I feel like, you can think what you want to think about me – that's OK. I have the most amazing fans, friends, and family members who have stood by me, and I know that my cast and crew really enjoy working with me. That's all that matters."


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Friday Five: Nobody
This Friday Five feature on The 9513 is all about nobody. You know the one. Nobody in particular. Nobody knows the trouble I've seen. That nobody. On the surface, you might think it might be difficult to find five songs about nobody. But in this case, nobody is a somebody and he's had thousands of songs written about him. So here, I am, trying to tell everybody–anybody that will listen–about these five songs about nobody.
- 5. "Nobody" – Johnny Cash
Cash says "I ain't never done nothin' to nobody, I ain't never got nothin' from nobody, no time, and until I get something from somebody, sometime, I don't intend to do nothin' for nobody, no time." Well that's clears things up, don't it?
- 4. "Nobody But You" – Dolly Parton
For decades, Dolly Parton used her sex appeal along with her amazing God-given talent to win over audiences on stage, in stores and on screen. It turns out that when her "lips are burning with desire" and when "only your kisses can put out the fire," the man to do it is, you guessed it, that nobody feller again. Lucky devil.
- 3. "Nobody Loves You Like I Do" – Patty Loveless
If we've already assessed that nobody is actually a somebody, does that mean that this song isn't as much a devotional pledge as it is a realization that the song's subject is in love with a second woman? How very country music of Patty.
- 2. "Nobody Drinks Alone" – Keith Urban
In one of Urban's best career tracks, this haunting song tells how nobody really drinks alone. He drinks along with all of your demons, your ghosts from your past and every memory you've held back. That's quite the dinner party.
- 1. "Nobody's Lonesome for Me" – Kris Kristofferson & Hank Williams
Two country legends perform this old Hank Sr. classic where everybody is getting all the glory and nobody gets the shaft. It has one of the better country music lines: "When the time rolls around for me to lay down and die, I bet I'll have to go and hire me someone to cry, Everybody's lonesome for somebody else, nobody is lonesome for me." Count me among the nobodies.
26 Comments
And of course "Nobody", by Sylvia.
Patty Loveless!
There is a fun video from the Glen Campbell Goodtime Hour featuring Glen along with Jerry Reed and Jackie DeShannon singing "Ain't Nobobdy's Business But My Own". Gosh did I love to watch that show when it was on the air…
Link: http://www.youtube.com/watch?v=8EdS_Vuz65s
Quick Opry Alert! Tonight's show will include Ralph Stanley, Josh Turner, and Kenny Rogers (that's what I call covering a lot of bases), and the Brock McGuire Band(?). Saturday night's Opry features Terri Clark, Diamond Rio and Troy Olsen.
Opry Schedule: http://www.opry.com/shows/ThisWeek.html
Makes me think of the Shel Silverstein poem about how "nobody loves me, nobody cares, nobody brings me peaches and pears," and so on, listing all the things "nobody" does for him, and then one day, "nobody wasn't there!"
"Nobody Loves Me," written by Zeke Clements, ought to be on that list – on the top.
"Nobody Love, Nobody Gets Hurt", the title song from the 1998 Suzy Bogguss album. The song was solely written by Bobbie Cryner.
"Nobody But Me" by Blake, his first sinigle after the mullet.
"Nobody But a Fool" – recorded by Connie Smith and a few more including an early Chely Wright. Written by the great Bill Anderson.
Like a few previous commenters, I also thought of Sylvia's "Nobody" and Blake's "Nobody But Me." But it's nice to see that great old Patty song on this list – one that I've always loved. Good ones from Johnny and Dolly too.
Might also point out that Sylvia's song is one in which Nobody really is an actual person.
I think that's what I always loved most about that song Ben. It's one of those 'slap you in your face before you see it coming' kind of songs/hooks.
"Nobody in His Right Mind Would've Left Her" – George Strait, written by Dean Dillon.
Sylvia's Nobody is my personal fav!
Oh my god, I'm so glad "Nobody Drinks Alone" is on the list. I think that is one of the most powerful songs ever recorded in country. So good.
Whoa, I'd never heard Nobody Drinks Alone… damn, Keith.
I've always been partial to the "Nobody Knows" taken from R&B singer Tony Rich… I'm not even a big fan of Kevin Sharp's vocal, but it's just a classic song, regardless of genre/artist.
Conversely, I've never really liked "Nobody But Me". Super catchy, but when you listen to the lyrics, it's kind of bizarre. "Nobody but me's gonna love you like you oughta be loved on, nobody but me's gonna cry if you ever leave." Basically, "I'm the best you're going to get, because no one else really cares about you". Touching.
The first one I thought of was Radney Foster's "Nobody Wins".
Happy Friday! :)
The first one that came to mind for me from country was Reckless Kelly's "Nobody's Girl". From the folk-pop world, The Weepies' "Nobody Knows Me At All" (it's actually pretty country-leaning. "Nobody Girl" by Ryan Adams also comes to mind.
oops, i meant to put a closing parenthesees after "country-leaning"
"Nobody Knows But Me" written and recorded by Jimmie Rodgers on November 25th 1929… and subsequently recorded by Lefty Frizzell, Hank Snow and Merle Haggard.
Also, "Nobody's Child" by Hank Snow on a late fifties lp called "When Tragedy Strikes."
"Nobody Wins" written and recorded by Kris Kristofferson.
SYLVIA's "Nobody" One of my favorite songs from the 80s…I still smile when I hear it on Sirius or on the rare occassion I hear it on regular radio
Nice call on "Nobody's Child," Patrick – can't believe I forgot it, since my band recorded it on our first album…
Brenda Lee had a great recording on KK's "Nobody Wins"
How did we miss the old spiritual "Nobody Knows The Trouble I've Seen", recorded by Louis Armstrong, Marian Anderson, Lena Horne and many of the 60s folkies. I especially surprised Jon didn't mention it
"Nobody Loves Me Like You Do" the Anne Murray/Dave Loggins #1 duet from December 1984
A few more:
Clint Black's "Nobody's Home"
Kathy Mattea's "Nobody's Gonna Rain on Our Parade"
Wynonna's "Can't Nobody Love You (Like I Can)"
George Strait's "Nobody In His Right Mind Would Have Left Her"
Joe Nichol's "If Nobody Believed in You"
and Ronnie Milsap's "Nobody Likes Sad Songs"
and a some from the pop world:
Carly Simon's "Nobody Does It Better"
Christina Aguilera & Ricky Martin's "Nobody Wants to Be Lonely"
and Madonna's "Nobody Knows Me"
First thing that came to mind was the lyrics to "When I Call Your Name" by Vince Gill. . . until I remembered the title– it doesn't mention nobody.
Nobody Drinks Alone is a dang good song!
Current Discussion
- Dave D.: I never expected to see a shout out to the Cameron Family Singers today (or any day, for that matter). …
- nm: Duncan Cooper is not wrong.
- bob: Re Herndon, O'Neal & Griggs touring together, Herndon's "Lies I Told Myself Album" includes a duet with O'Neal, "I Can't". …
- luckyoldsun: Looks like CMT is pulling a "surprise" for its Artists' Choice countdown. They're coming up on #2 but there are …
- Jim Z: " Luke Bryan (Best Country Male Artist), Brantley Gilbert's Just as I Am (Best Country Album), Carrie Underwood (Best Country …
- luckyoldsun: MH-- "...divisive, dismissive and argumentative." There's a shock. Who'da thunk it.
- MH: Looks like the IBMA is finding out what a lot of us already knew about former & frequent Engine 145 …
- Tom: ...it is almost the year 2015 - ain't it about time we leave people (celebrities included) having the personal relations …
- Paul W Dennis: I had assumed that Herndon was gay after the incident that Lucky Old Sun referenced, but bought the next Herndon …
- Jonathan Pappalardo: For anyone who wants to buy it, the Rosanne Cash 45 is available again. With 1000 copies being created, I …


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Blaine (4-1) kept pace with Elk River at the top of the Northwest Suburban Conference North Division after a 27-11 win at Andover (1-4) Sept. 26.
tom develice
Blaine (3-1) celebrated homecoming in style with a 35-3 win to keep Coon Rapids (0-4) searching for its first win of the season in a North Division of the Northwest Suburban Conference contest.
A wild comeback by Blaine in its home opener Friday gave the fans dressed in patriotic gear a reason to rush the field after pulling off a 22-21 win against rival Anoka.
Senior Ben Beckman and junior Griffin Fussy will compete for the starting quarterback spot after graduating longtime quarterback Alex Copa last season. Develice said the QBs are similar in style, having plenty of success during the baseball season. He likes the strength from Beckman, realizing Fussy isn't far behind.
Unlike their regular season meeting in which 62 points were scored two weeks earlier, the Section 5-6A semifinal between Blaine (6-3) and Centennial (5-4) favored the defenses with the Cougars earning a 17-11 win at Blaine Oct. 25.
Goodrich Field was transformed into Dave Brock Field for the annual Pumpkin Bowl football game, but Blaine walked away with a 21-9 win over Anoka, Oct. 16.
Blaine won its third straight game and posted a season-high points total in a 42-20 win at Centennial Oct. 11. The Bengals (5-2) have things going the right direction after back-to-back losses to good Osseo and Maple Grove programs in weeks three and four.
Blaine (4-2) overcame five turnovers and quarterback Alex Copa ran for 235 yards and two touchdowns including a four-yard run in overtime as the Bengals topped Champlin Park 21-14 Oct. 4.
Blaine put two losses behind them while spoiling Coon Rapids' homecoming game 41-14, Sept. 27. Cardinals Stadium was hoping with a rabid student section proud to share their Cardinal Nation pride throughout the deceptively one-side score.


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Why Leftism, though secular, must be understood as a religion.
Ronald Kessler
December 10th, 2014
Los Angeles, CA
Register Here
Bret Stephens
January 23rd, 2015
Los Angeles, CA
Register Here
March 6-8, 2015
Terrenea Resort
Palos Verdes, CA
Anthea Butler: google.com/search?q=anthe…
Black racist Ivy League prof sounds off: campusreform.org/?ID=6102&advD=…
Michael Brown's father last night: "Burn this bitch down." Obviously Michael was a chip off the old block.


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- 2 Slice cool-touch toaster
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Love the idea of this. Don't love the result. Inconsistent toasting, and only able to get the steelers toasted onto white bread.
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