text:
Serendipitous discovery and X-ray structure of a human phosphate binding apolipoprotein.
ABSTRACT We report the serendipitous discovery of a human plasma phosphate binding protein (HPBP). This 38 kDa protein is copurified with the enzyme paraoxonase. Its X-ray structure is similar to the prokaryotic phosphate solute binding proteins (SBPs) associated with ATP binding cassette transmembrane transporters, though phosphate-SBPs have never been characterized or predicted from nucleic acid databases in eukaryotes. However, HPBP belongs to the family of ubiquitous eukaryotic proteins named DING, meaning that phosphate-SBPs are also widespread in eukaryotes. The systematic absence of complete genes for eukaryotic phosphate-SBP from databases is intriguing, but the astonishing 90% sequence conservation between genes belonging to evolutionary distant species suggests that the corresponding proteins play an important function. HPBP is the only known transporter capable of binding phosphate ions in human plasma and may become a new predictor of or a potential therapeutic agent for phosphate-related diseases such as atherosclerosis.
- SourceAvailable from: Carlos Contreras-Martel[Show abstract] [Hide abstract]
ABSTRACT: We report the serendipitous discovery of a human plasma phosphate binding protein (HPBP). This 38 kDa protein is co-purified with paraoxonase (PON1). The association between HPON1 and HPBP is modulated by phosphate and calcium concentrations. The HPBP X-ray structure solved at 1.9 A resolution is similar to the prokaryotic phosphate solute-binding proteins (SBPs) associated with ATP binding cassette transmembrane transporters, though phosphate-SBPs have never been characterized or predicted from nucleic acid databases in eukaryotes. However, HPBP belongs to the family of ubiquitous eukaryotic proteins named DING, meaning that phosphate-SBPs are also widespread in eukaryotes. The absence of complete genes for eukaryotic phosphate-SBP from databases is intriguing, but the astonishing 90% sequence conservation of genes between evolutionary distant species suggests that the corresponding proteins play an important function. HPBP is the first identified transporter capable of binding phosphate ions in human plasma. Thus it is thought to become a new predictor and a potential therapeutic agent for phosphate-related diseases such as atherosclerosis.Annales Pharmaceutiques Françaises 04/2007; 65(2):98-107.
Article: A high throughput serum paraoxonase assay for discovery of small molecule modulators of PON1 activity.[Show abstract] [Hide abstract]
ABSTRACT: PON1 has been demonstrated to be the serum enzyme responsible for detoxifying organophosphate chemical weapons and plays a protective role against atherosclerosis. In order to identify small molecules that modulate PON1 activity in serum, we developed a high throughput kinetic absorbance assay using mouse serum and the organophosphate paraoxon. The IC(50) value obtained for the known PON1 inhibitor, 2-hydroxyquinoline, matched the value reported for purified PON1. A compound library was screened resulting in no confirmed activators, but 12 confirmed inhibitors. Seven of these hits also inhibited purified human PON1. One compound was only two-fold less potent than 2-hydroxyquinoline in the serum assay, but 10-fold more potent against purified PON1. This compound (IC(50) = 420 nM) may be useful towards a chemical probe for PON1. Therefore, this assay has utility as a high throughput assay for discovery of small molecule modulators of PON1 activity that maintain activity in serum.Current Chemical Genomics 01/2008; 2:51-61.
- [Show abstract] [Hide abstract]
ABSTRACT: DING proteins constitute an intriguing family of phosphate-binding proteins that was identified in a wide range of organisms, from prokaryotes and archae to eukaryotes. Despite their seemingly ubiquitous occurrence in eukaryotes, their encoding genes are missing from sequenced genomes. Such a lack has considerably hampered functional studies. In humans, these proteins have been related to several diseases, like atherosclerosis, kidney stones, inflammation processes and HIV inhibition. The human phosphate binding protein is a human representative of the DING family that was serendipitously discovered from human plasma. An original approach was developed to determine ab initio the complete and exact sequence of this 38 kDa protein by utilizing mass spectrometry and X-ray data in tandem. Taking advantage of this first complete eukaryotic DING sequence, a immunohistochemistry study was undertaken to check the presence of DING proteins in various mice tissues, revealing that these proteins are widely expressed. Finally, the structure of a bacterial representative from Pseudomonas fluorescens was solved at sub-angstrom resolution, allowing the molecular mechanism of the phosphate binding in these high-affinity proteins to be elucidated.Journal of Synchrotron Radiation 01/2011; 18(1):45-9. · 2.19 Impact Factor
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Structure 14, 601–609, March 2006 ª2006 Elsevier Ltd All rights reservedDOI 10.1016/j.str.2005.12.012
Serendipitous Discovery and X-Ray Structure
of a Human Phosphate Binding Apolipoprotein
Renaud Morales,1Anne Berna,2Philippe Carpentier,1
Carlos Contreras-Martel,1Fre ´de ´rique Renault,3
Murielle Nicodeme,4Marie-Laure Chesne-Seck,6
Franc ¸ois Bernier,2Je ´ro ˆme Dupuy,1
Christine Schaeffer,7He ´le `ne Diemer,7
Alain Van-Dorsselaer,7Juan C. Fontecilla-Camps,1
Patrick Masson,3Daniel Rochu,3
and Eric Chabriere3,5,*
1Laboratoire de Cristallogene `se et Cristallographie
des Prote ´ines
Institut de Biologie Structurale JP EBEL
38027 Grenoble
2Institut de Biologie Mole ´culaire des Plantes du CNRS
Universite ´ Louis Pasteur
67083 Strasbourg Cedex
3Unite ´ d'Enzymologie
De ´partement de Toxicologie
Centre de Recherches du Service de Sante ´ des Arme ´es
38702 La Tronche
4Laboratoire des Biosciences de l'Aliment
5Laboratoire de Cristallographie et Mode ´lisation des
Mate ´riaux Mine ´raux et Biologiques
Universite ´ Henri Poincare ´-Nancy 1
54506 Vandoeuvre-le `s-Nancy
6Laboratoire de Cristallographie Macromole ´culaire
Institut de Biologie Structurale JP EBEL
38027 Grenoble
7LaboratoiredeSpectrome ´triedeMasseBio-Organique
67087 Strasbourg
We report the serendipitous discovery of a human
plasma phosphate binding protein (HPBP). This 38
kDa protein is copurified with the enzyme paraoxo-
nase. Its X-ray structure is similar to the prokaryotic
phosphate solute binding proteins (SBPs) associated
with ATP binding cassette transmembrane trans-
porters, though phosphate-SBPs have never been
characterized or predicted from nucleic acid data-
bases in eukaryotes. However, HPBP belongs to the
family of ubiquitous eukaryotic proteins named
DING, meaning that phosphate-SBPs are also wide-
spread in eukaryotes. The systematic absence of com-
plete genes for eukaryotic phosphate-SBP from data-
basesis intriguing,but
sequence conservation between genes belonging to
evolutionary distant species suggests that the corre-
sponding proteins play an important function. HPBP
the astonishing90%
phate ions in human plasma and may become a new
predictor of or a potential therapeutic agent for phos-
phate-related diseases such as atherosclerosis.
Both cholesterol and HDL (high-density lipoprotein)
levels are commonly used as risk predictors of cardio-
vascular disease (Gordon and Rifkind, 1989; Amann
et al., 2003). HDL protects against atherosclerosis
mainly through the reverse cholesterol transport pro-
cess in which excess cholesterol is transferred from
peripheral tissues to the liver by the ATP binding cas-
sette ABCA1 transporter, which is defective in Tangier
disease (Brooks-Wilson et al., 1999). Although HDLs
have been extensively studied, their metabolic role
has not been completely elucidated yet. For instance,
while the involvement of the calcium-dependent, HDL-
associated paraoxonase 1 (PON1; aryldialkylphospha-
tase; EC 126.96.36.199) in atherosclerosis prevention is well
established (Watson et al., 1995; Shih et al., 1998), its
physiological function remains unknown. PON1, named
for its ability to hydrolyze the insecticide paraoxon, is
the subject of intensive research owing to its capacity
to inactivate various organophosphorous compounds,
including nerve gases and pesticides, representing both
a terrorist threat and an environmental hazard. Albeit its
native activity is suggested to be lactonase (Khersonsky
and Tawfik, 2005), PON1 is currently considered as an
enzyme with promiscuous functions. Accordingly, some
of the activities attributed to PON1 are for the moment
largely debated.
In preparing PON1 for functional studies, we have iso-
lated another protein that copurifies with it. Here, we re-
port the discovery and X-ray structure of this protein,
which is a human plasma solute binding protein (SBP)
that binds phosphate and belongs to the sixth family
of SBPs (Tam and Saier, 1993). In prokaryotes, SBPs
areassociatedwithABCtransporters, afamilyofubiqui-
tous molecules that use ATP as an energy source by
which to carry a wide variety of molecules across mem-
branes (Higgins, 1992). ABC transporters may also play
a central role in pathologies such as cystic fibrosis and
are involved in resistance to chemotherapeutic drugs
(Jones and George, 2002). These proteins are com-
posed of two cytoplasmic nucleotide binding domains
and two channel-forming transmembrane domains
(Chang and Roth, 2001). In both archaea and bacteria,
interactions with extracytoplasmic SBPs largely con-
tribute to their efficiency and specificity. Prokaryotic
phosphate-SBPs are 38 kDa proteins, encoded by
PstS genes (Surin et al., 1984), that bind inorganic phos-
phate ions with submicromolar affinity (Medveczky and
Rosenberg, 1971) and are mostly expressed during
phosphate starvation (Gerdes and Rosenberg, 1974).
In gram-negative bacteria, these proteins are soluble
and periplasmic, whereas in gram-positive bacteria,
they are lipoproteins anchored to the outer face of the
membrane (Young and Garbe, 1991).
*Correspondence: firstname.lastname@example.com
Page 2
Interestingly, eukaryotic ABC transporters do not rely
on SBPs. Proteins related to SBPs nevertheless exist in
eukaryotes (Felder et al., 1999), the more relevant cases
being the glutamate receptors, the atrial natriuretic pep-
tide receptors involved in blood pressure regulation,
and the calcium-sensing G protein-coupled receptors,
located in parathyroid and kidney cells. Anchored to
the membrane with an extramembranar domain, they
act in a multimeric state as receptors.
This article describes the discovery, characterization,
sequencing, and crystal structure at 1.9 A˚resolution of
HPBP, a novel, to our knowledge, human HDL-associ-
ated apolipoprotein, not predicted by any genomic da-
tabase.Thisprotein,structurally andfunctionally related
to bacterial phosphate-SBPs, may be the first phos-
phate transporter characterized in human plasma. A
possible use of the protein in diagnosis or therapy of
atherosclerosis emerges from its capability of binding
phosphate, as well as a role for stabilizing functionally
active conformation of PON1.
X-Ray Structure of HPBP
Crystals were obtained from an apparently pure PON1
solution (Contreras-Martel et al., 2006; see the Supple-
mental Data, available with this article online). Due toini-
tial difficulties in obtaining heavy atom derivatives, the
protein envelope was initially determined ab initio at
25 A˚resolution (Fokine et al., 2003). Eventually, a uranyl
at 1.9 A˚resolution and a polyalanine model was traced
automatically into this map (Figures 1A and 1B) (see Ex-
perimental Procedures). As the electron density did not
match the amino acid sequence of PON1, we carried out
structural comparisons with DALI (Holm and Sander,
1994). This analysis revealed that our three-dimensional
model (PDB ID code 2cap) displayed a fold very similar
to those of a phosphate binding protein from Escheri-
chia coli (Luecke and Quiocho, 1990) and the PstS1 pro-
tein from Mycobacterium tuberculosis, a major BCG
vaccine antigen (Vyas et al., 2003). Based on these ob-
servations, the protein was named human phosphate
binding protein (HPBP). The complete amino acid se-
quence ofHPBP was assigned from the electron density
map and later confirmed for about 75% of the protein by
direct sequencing (Figure 2 and Experimental Proce-
dures). HPBP consists of 376 residues with a predicted
molecular mass of 38.4 kDa.
Superposition of the three-dimensional structures
gave rms deviations for a carbon atom positions be-
tween HPBP and E. coli phosphate-SBP (216 atoms)
Figure 1. HPBP Structure
(A) Ribbon representation of HPBP showing
the two domains (blue and green) connected
by a hinge (yellow), the phosphate molecule
(red balls), and the two disulfide bridges
(C113–C158 and C306–C359, orange sticks).
The elongated protein structure exhibits two
similar globular domains, each constituted
by a five-stranded b sheet core (three parallel
strands followed
b strands) flanked by a helices. A hydropathy
profile computed with the program DAS
(Cserzo et al., 1997) indicates that HPBP is
strongly hydrophobic between residues 168
and 194. This motif, located on the protein
surface (H.S.), is involved in the packing of
the crystallographicdimer. N and C represent
the protein termini.
(B) Stereoview of the Ca trace of HPBP. The
orientation is the same as in (A). Every tenth
a-carbon is labeled with its residue number.
Rainbow coloring—from the N terminus to
the C terminus—has been used.
(C) Structural comparison of different known
phophate-SBPs: HPBP is shown in blue, E.
coli PstS protein is shown in yellow, and M.
tuberculosis PstS1 is shown in green. The
four HPBP-specific loops are indicated.
by twoantiparallel
602 602
Page 3
and HPBP and M. tuberculosis PstS1 protein (202
atoms) of 1.53 A˚and 1.52 A˚, respectively. Four loops
protrudring from HPBP globular domains represent its
only major structural difference with bacterial SBPs
(Figure 1C).
An HPBP electron density map displays a peak that is
completely buried in a cleft formed at the intersection of
the two domains and that could be assigned to either
a phosphate (HPO422) or a sulfate (SO422) ion (Fig-
ure 3A).
Ligand Binding
This active site is very similar to other phosphate-SBPs.
Over the 8 residues involved in the binding of the sub-
strate, F11 in E. coli PBP is replaced by L9 in HPBP
(Figure 3B). Moreover, this change has only a minor ef-
fect because this residue binds the substrate with its
NH backbone. The dibasic phosphate is bound by
a rich network comprising 13 hydrogen bonds. Twelve
of them involve dipolar donor groups with six backbone
NHs (T8, L9, S32, S144, G145, T146), mostly located at
the ends of three helices; two NH side chains from
R114; and four from side chains OHs (T8, S32, S144,
T146). R114 makes two hydrogen bonds with both neg-
ative oxygen atoms of the dibasic phosphate. The pos-
itive charge of R114 is neutralized by a salt bridge with
E142. This interaction, by diminishing the charge cou-
pling interaction between the guanidium and phos-
phate, has previously been described as essential for
the fast release of the ionic substrate bound with
charged residues (Vyas et al., 2003). D61, the only dipo-
lar acceptor group, makes a hydrogen bond with the
only proton available on dibasic phosphate. This key
residue is responsible for the discrimination by five or-
ders of magnitude between HPO422and the closely re-
lated sulfate (Luecke and Quiocho, 1990). Sulfate at
physiological pH does not possess any proton and will
be repelled by the aspartic residue. This hydrogen
Figure 2. Sequence Conservation between HPBP and SBPs or DING Proteins
Alignment of the amino acid sequence of HPBP deduced from the crystal structure (residues confirmed by direct amino acid sequencing are in
bold characters) with various SBPs and DING proteins by using the program ALIGN (Myers and Miller, 1988) Potato, potato DING protein (A.B.
et al., unpublished data); L. major, L. major Friedlin almost complete DING protein sequence translated from contigs 2206 and 2072 from the L.
major Friedlin shotgun sequencing program; CAI, human CAI N-terminal and internal peptide fragments (Kumar et al., 2004); E. coli PstS (Gen-
Bank AAA24378.1); and M. tuberculosis PstS1 (GenBank M30046). Boxes indicate conserved residues; arrows indicate residues involved in
phosphate binding in E. coli and M. tuberculosis PstS proteins. The position of protruding loops is indicated. * and # indicate cysteines involved
in disulfide bridges.
A Human Phosphate Binding Apolipoprotein
Page 4
bond in our structure is short (2.44 A˚) (Figure 3B), as de-
scribed in other phosphate-SBPs at ultra high 0.98 A˚
resolution. Despite this short distance, this hydrogen
bond would not be stronger than other hydrogen bonds
(Wang et al., 1997). Finally, the terminal oxygen atom is
bound with hydrogen donors. According to the struc-
ture, the binding of the monobasic phosphate could be
done similarly to other phosphate-SBPs without any
drastic rearrangement. The g-OHof S32is the only other
groupfavorably positioned toacceptthrough its oxygen
lonepair the second phosphate proton (on O3) while do-
nating its proton to the D61. The negative charge of the
phosphate will be concentrated on the two oxygen
atoms (O1 and O2) (Luecke and Quiocho, 1990). In the
case of HPBP, the surface of the binding pocket is neg-
ative, as described in other phosphate-SBP and sulfate-
SBPs (Ledvina et al., 1996; Vyas et al., 2003). It has been
proposed that this peculiar feature of an anion bound
within a negative environment would increase the dis-
crimination between negative substrates. In this way,
only anions perfectly matching the hydrogen bond net-
vorable electrostatic interaction of the anion.
The binding of dibasic phosphate has been experi-
mentally determinedby32P-orthophosphate bindingas-
says (Figures 3C and 3D). Dialyses experiments (see Ex-
phosphate at pH 8 with a better than submicromolar af-
finity, as described for other phosphate-SBP and DING
proteins (Scott and Wu, 2005). Although, monobasic
phosphate binding has been observed for bacterial
phosphate-SBPs, no phosphate binding has been ob-
served at pH 4.5 for HPBP. Small differences at the
active site between HPBP and these of bacterial
that HPBP binds
Figure 3. Binding of the Phosphate Ion
(A) Stereoview of the binding site. Residues involved in the binding of the phosphate molecule are labeled.
(B) Schematic representation of the hydrogen bond network involved in phosphate binding. Amino acids involved in phosphate binding in both
HPBP and E. coli PstS (in italics) are indicated. Numbers represent hydrogen bonds lengths in A˚.
(C) In vitro HPBP phosphate binding. PON1/HPBP were purified from six plasma samples (lanes a–f) and assayed for phosphate binding with
radiolabeled phosphate (see Experimental Procedures). Lanes g and h correspond to pure b-lactoglobulin and lysosyme, respectively, used
as negative controls.
(D) Two different PON1/HPBP-containing fractions purified from two different plasma bags (lanes a and b) separated by SDS-PAGE. Proteins
were revealed by Coomassie staining (left) and phosphate binding by autoradiography (right). PON1 and HPBP have been identified by N-ter-
minal amino acid sequencing.
Page 5
phosphate-SBPs could not explain this fact. Moreover,
no binding has been observed after redialysis at pH 8.
Therefore, it is likely that that HPBP is denatured at pH
4.5, a nonphysiological environment for a plasma pro-
Association of HPBP with PON1
Initial efforts to separate PON1 from HPBP by DEAE
chromatography failed even though the two proteins
have different pIs (Figure 4). The very harsh conditions
required to separate HPBP from PON1 by using two-
dimensional PAGE (see Experimental Procedures) sug-
gest that PON1 and HPBP copurified as a heteroo-
ligomer with two subunits of almost identical molecular
weights. Indeed, using analytical ultracentrifugation,
Josseetal.(2002)showed thatPON1(consideredas ho-
mogeneous at this moment) was oligomeric. Recent
studies (data not shown) obtained by gel filtration chro-
matography indicate that the oligomeric composition of
PON1, HPBP, and PON1/HPBP complex(es) are depen-
dent on the calcium, phosphate, and detergent concen-
HPBP crystallizes as a homodimer that we assume to
be physiologically relevant because its interface, which
is mostly hydrophobic, has an area of about 790 A˚2,
a typical value for transient protein complexes (Jones
and Thornton, 1998). The conformational changes in-
duced by the composition of the crystallization liquor
could have favored the dissociation of the complex
and privileged crystallization of HPBP alone. Indeed, it
has been shown that, upon substrate binding, SBPs un-
dergo a conformational change concerning the two do-
mains that are connected by a flexible hinge (Felder
et al., 1999). This movement, described as a ''Venus fly
trap motion'' (Mao et al., 1982; Yao et al., 1996), was
also predicted for HPBP by using the program ElNemo
(Suhre and Sanejouand, 2004) (see Movie S1 available
with the Supplemental Data online).
Eukaryotic Phosphate-SBPs Are Related to DING
Surprisingly, a gene sequence corresponding to HPBP
has not been found in either the human genome or
EST databases. Nevertheless, we have been able to re-
late HPBP to a family of still poorly described proteins
that have been named DING according to the sequence
of their 4 conserved N-terminal residues (Berna et al.,
2002; A.B. et al., unpublished data). Proteins belonging
to this family are ubiquitous, since they have been iden-
tified in animals (man, monkey, rat, turkey), plants (Ara-
bidopsis thaliana, potato, tobacco), and fungi (Candida
albicans) (Riah et al., 2000; Berna et al., 2002; Belenky
et al., 2003; Blass et al., 1999; Kumar et al., 2004; Adams
et al., 2002; Weebadda et al., 2001; Scott and Wu, 2005).
DING proteins are extracellular with a mass of w40 kDa.
As in the case of HPBP, which is associated with PON1,
several of the DING proteins are often purified as homo-
or heterooligomers. Closely related proteinsalsoexist in
some Pseudomonas strains (A.B. et al., unpublished
data). In addition, hundreds of other prokayotic proteins
display a lower, but significant, homology with eukary-
otic DING proteins, although they do not start with the
DING peptide. These proteins are the SBPs involved in
phosphate uptake together with ABC transporters (see
The 8residues involved in phosphate binding are con-
served in all prokaryotic SBPs and also in all eukaryotic
DING proteins (Figure 2 and A.B. et al., unpublished
data). Figure 2 also shows that the cysteines that
make two disulfide bridges in HPBP are probably also
conserved in all eukaryotic DING proteins as well as in
most prokaryotic proteins related to them.
HPBP displays about 64% sequence identity with
DING proteins from potato and Leishmania major,
whereas HPBP sequence identities with E. coli and M.
tuberculosis phosphate-SBPs display only 25% and
23%, respectively (Figure 2).
HPBP Biochemical Properties
HPBP was found to be tightly associated with PON1.
Besides, subsequently to our description of a copurified
protein in supposedly pure PON1 preparations (Fokine
et al., 2003), recent studies indicate that the presence
of additional proteins in purified PON1 fractions is not
uncommon and could lead to confusion in the interpre-
tation of PON1 function (Teiber et al., 2004; Connelly
et al., 2005; Draganov et al., 2005). In at least one case,
a protein copurified with PON1 had a molecular weight
and a pI corresponding to HPBP (Smolen et al., 1991).
PON1 is an apolipoprotein largely located on HDL but
also present on chylomicrons and VLDL (Fuhrman
et al., 2005; Deakin et al., 2005). Thus, we assume that
the previously unknown hydrophobic HPBP is an apoli-
poprotein associated in vivo with PON1.
Moreover, experiments assessing the characteriza-
tion of the oligomeric organization of the PON1/HPBP
as well as its impact on the functional activity of the
two proteins, which both appear as dependent to cal-
cium, phosphate, and detergent (in vitro) or lipoproteins
(in vivo) amounts, are in progress (M. Elias et al., per-
sonal communication).
Figure 4. Two-Dimensional PAGE of the Purified PON1 Fraction
Identification by two-dimensional PAGE of proteins in a purified
PON1 fraction obtained according to Gan et al. (1991). The first di-
mension corresponds to isoelectric focusing in a 3–10 pH gradient;
the second dimension is 10% SDS-PAGE. PON1 and HPBP, identi-
fied by N-terminal sequencing, have similar masses (w40 kDa), but
very different pIs (w5 and w8, respectively). The right lane shows
molecular weight markers.
A Human Phosphate Binding Apolipoprotein
Page 6
HPBP structure clearly relates it to bacterial phos-
phate-SBPs associated with ABC transporters. We
have shown that HPBP is capable of binding inorganic
phosphate ions at physiological conditions with submi-
cromolar affinity. The normal concentration of free inor-
ganic phosphate in human plasma is about 0.5 mM; ow-
ing to its high specificity, HPBP is always saturated in
phosphate.For now,wecanonlyspeculate on thephys-
iological function of this first, to our knowledge, phos-
phate binding protein reported so far in human plasma
(see below).
Eukaryotic Phosphate-SBPs Are Missing from
knowledge, member of the DING family of proteins.
Even though DING proteins are likely ubiquitous in eu-
karyotes, they remain poorly characterized since very
few coding sequences are present in DNA or RNA data-
bases and since none of them is complete (A.B. et al.,
unpublished data). Thus, HPBP may provide the first
complete sequence of a DING protein. To our knowl-
edge, recent cloning of a large part of the potato DING
gene and discovery of two partial DNA sequences of
DING proteins in unannotated parts of the Leischmania
major and Rattus norvegicus genomes allowed for ex-
tensive comparison between different DING genes for
the first time (A.B. et al., unpublished data). This re-
vealed an astonishing conservation among several of
the DING proteins and genes, with pairwise identities
above 95% and 90% at the amino acid and nucleotide
levels, respectively.
The very high conservation of some phosphate-SBP
gene sequences between distant species such as po-
tato (a higher plant) and Leishmania major (a protozoan)
cannot be explained by a constraint based only on
a physiological function requiring the integrity of the
protein. These observations should stimulate studies
to determine unknown features that could prevent iden-
tification of some specific genes in systematic sequenc-
ing approaches.
In humans, DING proteins have previously been iso-
vial stimulatory protein that could be involved in the de-
velopment of rheumatoid arthritis (Hain et al., 1996;
Blass et al., 1999). Subsequently, a related protein se-
creted from fibroblasts and cervical carcinoma cells
was purified by virtue of its affinity for hirudin (Adams
et al., 2002). A protein displaying high affinity for genis-
tein, an estrogen analog, was identified in breast cells
(Belenky et al., 2003). Finally, a protein secreted by kid-
ney epithelial cells was named crystal adhesion inhibitor
(CAI) (Kumar et al., 2004). With a total of 261 amino acids
obtained by N-terminal and internal peptide sequenc-
ing, CAI was, up until now, the longest DING protein se-
quence known. At 59% amino acid identity with CAI,
HPBP clearly belongs to a different DING protein sub-
family. Thus, at least two DING genes are missing from
the sequenced human genome.
By possibly providing the first structure and the first
lated DING proteins and bacterial phosphate-SBPs.
This conclusively demonstrates that this last class of
proteins is likely ubiquitous in living organisms, though
it has never been characterized or predicted from ge-
nome analyses in eukaryotes. In addition, since the 8
residues that constitute the phosphate binding site in
prokaryotic SBPs are conserved in DING proteins, it is
reasonable to assume that functional phosphate-SBPs
are present in all eukaryotes. However, their association
with ABC transporters remains to be characterized.
Physiological Function of Human DING Protein
The presence of an inorganic phosphate scavenger or
receptor in human plasma associated with HDL may
be explained by the need to prevent the interaction of
this anion with Ca2+. Indeed, hyperphosphatemia is
a cardiovascular disease risk factor (Amann et al.,
2003) because the insoluble calcium phosphate, along
with cholesterol, increases the formation of atheroscle-
rotic plaques that block blood vessels (Dorozhkin and
Epple, 2002). Interestingly, the related CAI protein has
been described to prevent the formation of kidney
stone, by inhibiting calcium phosphate precipitation
(Kumar et al., 2004). In plasma, HPBP should be always
saturated with phosphate; therefore, we could suppose
that HPBP could sense or transport phosphate in a low-
phosphate toward a specific acceptor, or, similarly to
the CAI, prevent the formation of calcium phosphate
Alloftheseconcomitant factssuggestthatdisorder of
HPBP might favor hyperphostemia and/or atherosclero-
sis plaque development. Thus, HPBP, serendipitously
discovered as tightly bound to the HDL lipoprotein
PON1, may contribute, with PON1, to the antiathero-
genicity of a category of HDL particles. Accordingly,
HPBP could be tentatively regarded as a new predictor
disorders, including atherosclerosis.
compartment, transport
Experimental Procedures
Crystallization and Heavy Atom Derivative Soaks
The HPBP/PON1-containing fractions were obtained as described
in the purification protocol (see the Supplemental Data). Fractions
were dialyzed and concentrated in the presence of C-12 maltoside
(0.64 mM) by using a centrifugation technique (Centriprep Amicon
cutoff: 10 kDa, Millipore, St. Quentin-en-Yvelines, France) to a final
O.D. of 2.3. Crystallization was performed by using the hanging
drop vapor diffusion method. A total of 3 ml protein solution was
mixed with 2 ml of a reservoir solution containing cacodylate buffer
(50 mM, pH 6), ammonium sulfate (2 M), and sodium chloride (1
mM). Smallcrystalsappeared after1weekat4ºC.Heavy atomderiv-
atives were obtained by soaking crystals during 3 days in a reservoir
solution containing a uranium salt at 0.1 mM (Uranium-Plasmocor-
inth B [Chesne, 2002], a generous gift from Olivier Bertrand).
Data Collection and Structure Determination
X-ray diffraction data were collected at 100 K by using synchrotron
radiation at the FIP BM30 beamline (ESRF, Grenoble, France) with
a MarCCD (165 mm) detector. A single data set was recorded at
a wavelength of 0.934 A˚from a native crystal, and two other data
sets were recorded at the uranium LIII edge from one derivatized
crystal (Table 1). X-ray diffraction data were integrated, scaled,
and merged with the XDS2000 program (Kabsch, 1993) and the
CCP4 program suite (CCP4, 1994). CNS (Brunger et al., 1998) and
SnB programs (Weeks and Miller, 1999) were used to find the posi-
tions of the three uranium atoms. Initial experimental phases were
obtained with the program SHARP (De la Fortelle and Bricogne,
1997) by using the single isomorphous replacement with anomalous
scattering (SIRAS) method with a figure of merit of 0.46. A solvent
Page 7
flattening procedure was applied to the electron density map by us-
ing SOLOMON (Abrahams and Leslie, 1996) and DM (Cowtan, 1994).
The polypeptide chain was partially built in an automatic mode by
using ARP/wARP (Perrakis et al., 2001). CNS was employed for the
structure refinement, and TURBO-FRODO (Roussel et al., 1990)
was used for manual model building. Statistics for refinement are
given in Table 1.
The complete amino acid sequence was first determined from X-ray
data. This was helpful for the alignment of sequences subsequently
obtained by N-terminal sequencing, mass spectrometry analysis,
and internal peptide digestion (Supplemental Data). About 75% of
the sequence has been confirmed, showing an error rate of only
11% in the crystallographically predicted sequence. So, the HPBP
sequence provided herein is complete and exact at 97%.
In Vitro Phosphate Binding Assays
For the dot-blot binding assays, HPBP/PON1-containing fractions
from different plasma samples and negative control proteins (lyso-
membrane. After incubation in 50 mM Tris (pH 8), 2 mM32P-ortho-
phosphate (10 mCi/ml) (Amersham) for 150 min, the membrane
wasrinsedtwice inthesamebufferwithout phosphate andwassub-
film. For the assays in gels, separation of HPBP and PON1 was ac-
complished by using SDS-PAGE under nonreducing conditions
(8-25 Phast gel Pharmacia) after preincubation of the HPBP/PON1-
containing fractions with 0.1% SDS without heating. The gel was
run and then rinsed four times for 15 min in 0.1% Triton X-100-con-
taining buffer to remove SDS, incubated with32P-orthophosphate,
and revealed as mentioned above or stained with Coomassie blue.
An equilibrium dialysis method was employed for studying bind-
ing of Pi to HPBP. Dialysis tubings with membrane with a molecular
weight cutoff of 8 kDa were filled with 400 ml of a solution containing
1 mM HPBP/PON1, 0.1% Triton (v/v), 150 mM NaCl, in 25 mM Tris-
HCl buffer (pH 8.0), or 25 mM acetate buffer (pH 4.5). Tubings were
immersed into vials containing the same solutions without protein,
supplemented with 0.01 mM, 0.1 mM, 1 mM, or 10 mM orthophosphate
labeled with32P-orthophosphate (10 mCi ml21) (Amersham). Dialy-
sis were performed during 1 week to allow achievement of equilib-
rium. Phosphate concentration in vials and dialysis tubings was cal-
culated from scintillation measurement.
Two-Dimensional Gel Electrophoresis
HPBP/PON1-containing fractions were prepared in 9.8 M urea, 4%
(v/v) Triton X-100, 2 mM tributyl phosphine, 0.2% of ampholytes
3–10 (Biolytes, Bio-Rad, Marnes-la-Coquette, France), and 0.001%
(m/v) bromophenol blue. A total of 40 mg protein was separated in
the first dimension by focusing in IPG-Strips (Bio-Rad) on a Protean
IEF cell (Bio-Rad) at 250 V for 15 min, voltage ramping to 4000 V for
2 hr, followed by focusing at 4000 V for 4 hr. Separated proteins of
the first dimension were resolved in the second dimension by stan-
dard SDS-PAGE. Gels were silver stained.
Supplemental Data
Supplemental Data including the protein purification protocol, ta-
bles of data collection and of refinement statistics, sequencing,
mass spectrometry analysis, and a movie showing the Venus fly
trap motion are available at http://www.structure.org/cgi/content/
This research was supported by grants to P.M., J.C.F.-C., and E.C.
by De ´le ´gation Ge ´ne ´rale pour l'Armement (CO nº010807/03-10); by
a grant to D.R. by Sanita ¨tsakademie der Bundeswehr, and by the
Centre National de la Recherche Scientifique.
Received: July 11, 2005
Revised: November 9, 2005
Accepted: December 20, 2005
Published online: March 14, 2006
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NativeU PeakU Remote
Wavelength (A˚)
Space group
Unit cell (A˚)
Number of observed
Number of unique
(last bin) (%)
Rsym(last bin) (%)
I/s(I) (last bin)
Last resolution shell
a = 97.4, b = 87.9, c = 90.5
328,312 129,426
30,19454,892 35,705
99.4 (91.8)98.8 (97.1)95.1 (93.5)
9.3 (23.6)
13.78 (7.31)
8.4 (29.7)
11.3 (3.7)
8.3 (16.0)
12.0 (6.3)
Refinement Statistics
Resolution range (A˚)
Number of reflections
used in refinement
Number of protein atoms
Number of water
Average B factor (A˚2)
Rms bond
length (A˚)
Rms bond
angle (º)
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ture factor amplitude and Fcdenotes the structure factor amplitude
calculated from the model. Rfreeis as for Rwork, but it is calculated
with 5% of the randomly chosen reflections omitted from the refine-
A Human Phosphate Binding Apolipoprotein
Page 8
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Page 9
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Accession Numbers
The atomic coordinates and structure factors of the HPBP structure
have been deposited with the Protein Data Bank with the accession
code 2cap.
A Human Phosphate Binding Apolipoprotein


text:
Chris Martin
Watch One Direction, Sam Smith, Ellie Goulding, Bono & More Perform 'Do They Know It's Christmas?' For Band Aid 30
Nearly every British artist ever gathered together this weekend to re-record the holiday classic, "Do They Know It's Christmas?" in honor of the charity single's 30th anniversary. That is, except for Adele.
Chris Martin is Back to Writing Songs for Rihanna
"She has so many amazing people writing for her. It's like trying to win American Idol, getting a song on Rihanna's album," Martin said.
Chris Martin Talks Coldplay's Plans And Studio Time With Rihanna
Coldplay's Chris Martin visited the Fresh studio for an exclusive interview with Cane and to play acoustic set (including hits "A Sky Full of Stars" and "Viva La Vida") for a few lucky fans.
Coldplay Frontman Chris Martin's Best Live Acoustic Performances
Check out some of Chris Martin's BEST acoustic performances of some of Coldplay's greatest hits…
Avicii Recruits Jon Bon Jovi, Chris Martin, Billie Joe Armstrong & More for Next Album
"It's going to be a lot more song-oriented," the DJ and producer explained. "True was an attempt at that, getting electronic music in a song format."
Coldplay's Chris Martin Talks New Album: 'It Isn't for Everybody'
Chris Martin doesn't think his band's new album is for everyone, but he's proud of it anyway.
Watch Coldplay Perform 'Always In My Head' On 'Tonight Show'
In front of dangling paper stars, Martin sang the rather somber song, which samples his daughter Apple's voice.
Rock Hall Induction 2014: Good Vibes, Special Guests and Lorde
The Rock and Roll Hall of Fame induction ceremony used to be a drunken, freewheeling event that would stretch to all hours of the night. Today it's an arena-rock spectacle with stunning moments that won't soon be forgotten.
Watch Coldplay Get 'Magic-al' in New Music Video
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Chris Martin and Gwyneth Paltrow Are Getting A Divorce
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subset: commoncrawl


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Cowboys coach Travis Ford said he liked his team's energy and devotion to defense and rebounding in a 79-45 rout of Texas Tech Saturday.
Oklahoma State basketball notebook: Travis Ford sends message of expectation
But he said, too, that it's only a start.
"No way around it, that's great for today," Ford said. "I told the team, 'This is who you need to be. This is who you should be. And this is the only thing we're going to accept. And, guys, if you're not going to do it, we've got other guys who are playing.
"'If you're not going to do it, you're going to sit over there. There's an approach. There's an attitude. There's a body language that we want to portray that we are. That's business for us right now. That's the way we're approaching it.'
"As young a team as we are, I feel like we've got to keep our thumb on that."
The effort came following a rugged week of work that featured several 6 a.m. practices, a response to the team's ragged effort in a loss at Oklahoma last Saturday.
One other thing Ford is looking to see — and he said it was there Saturday:
"They need to enjoy it," he said. "The look and glee in their eye on almost all of them, they were having fun playing hard. They were enjoying playing with that intensity. And that, as a coach, is fun to see.
"Consistency is the word."
One of OSU's lowlights of the 2012 season came at Baylor, where the Cowboys were beaten 106-65 in what went down as the Cowboys' second-worst defeat — ever — and the worst since 1932.
Heading back to Waco for a Monday clash with the Bears, Cowboys guard Markel Brown said there's nothing to glean from that game.
"We left that game behind," Brown said. "That was a terrible game we played. But it's time for us to go down there and respond and play the way we're capable of playing and kind of put some pressure on them."
OSU has lost 20 of its last 21 road games in the Big 12, yet needs to find a few breakthroughs to reestablish itself as a conference contender.
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495 Express Lanes free for one week during off-peak hours
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Md. woman relives painful childhood abuse by swim coach
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Russia sanctions put squeeze on area gun owners
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Repaving means business is slow on Connecticut Avenue
As road crews work around the clock to repave a stretch of Connecticut Avenue in Chevy Chase, area businesses are feeling the pain of the gridlock created by lane closures.
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New lines drawn for D.C.'s school system
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Pet lovers' pictures may give crooks the upper hand.
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Childhood hunger moving to the suburbs
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Hollywood Park Cancels Live Racing May 26
Pointing to an insufficient number of entries, Hollywood Park announced that it has canceled its May 26 program. Live racing will resume at the Inglewood, Calif., track on May 27.
Pointing to an insufficient number of entries, Hollywood Park announced that it has canceled its May 26 program. Live racing will resume at the Inglewood, Calif., track on May 27.
Newly arrived Make Music for Me had his first breeze at Belmont Park May 23, covering a mile in 1:43.65 with trainer Alexis Barba looking on in preparation for the June 5 Belmont Stakes (gr. I).
Monmouth Park followed its record-breaking opening day with another strong showing May 23. A total of $7,046,389 was wagered on the 11-race program, an increase of 126% over the corresponding day last year.
Trainer David LaCroix has decided to retire as a trainer, going out on a winning note at Hollywood Park May 19 when he saddled Drink at Buster's for a 22-1 upset in a maiden race for 2-year-olds.
Forever Together, the 2008 female grass champion, will ship west to take on Tuscan Evening, the current female star in California, in the $250,000 Gamely Stakes (gr. IT) at Hollywood Park May 29.
With a confident ride from Eurico Da Silva, heavily favored Biofuel registered her fourth win in five starts over Woodbine's Polytrack in the $158,370 La Lorgnette Stakes May 23.
Rapidly-improving Sarafina survived a stewards' inquiry to take the Montjeu Coolmore Prix Saint-Alary (Fr-I) by a half-length over Deluxe at Longchamp May 23.
Bethrah got up in the final strides to prevail by a head in a blanket finish in the Etihad Airways Irish One Thousand Guineas (Ire-I) at the Curragh May 23.
The first dead heat in a grade I race in Japanese history took place May 23 when favored Apapane and Saint Emilion battled to a draw in the $2.2 million Yushun Himba (Japanese Oaks) at a rainy and soggy Tokyo Racecourse.
Champion mare Goldikova made the perfect start to her 5-year-old campaign, winning the Prix d'Ispahan (Fr-I) by a half-length over Byword at Longchamp May 23.
Trainer: Art Sherman
Owner: Coburn, Steven and Martin, Perry
Jockey: Victor Espinoza
Trainer: Bob Baffert
Owner: Kaleem Shah, Inc.
Jockey: Martin Garcia
Trainer: Jerry Hollendorfer
Owner: Jungle Racing LLC, KMN Racing LLC, Hollendorfer, Litt, Solis II and Todaro
Jockey: Mike E. Smith
Trainer: Steven M. Asmussen
Owner: Winchell Thoroughbreds LLC
Jockey: Rosie Napravnik
Trainer: Christophe Clement
Owner: Robert S. Evans
Jockey: Joel Rosario
Trainer: H. Graham Motion
Owner: Flaxman Holdings, Ltd.
Jockey: John R. Velazquez
Trainer: Christophe Clement
Owner: Haras du Mezeray and Skymarc Farm, Inc.
Jockey: John R. Velazquez
Trainer: J. Keith Desormeaux
Owner: Brehm, Erich, Detmar, Wayne, Michaels, Lee, Desormeaux, J. Keith et al.
Jockey: Kent J. Desormeaux
Trainer: Josie Carroll
Owner: Melnyk Racing Stables, Inc.
Jockey: Luis Contreras
Trainer: D. Wayne Lukas
Owner: Willis D. Horton
Jockey: Paco Lopez


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Teaser Trailer for The Dark Knight Rises! »
'The Batman has to come back'
Former TV Batman Adam West to appear in Dark Knight Rises? »
Mayor West, is that you?
A Round-Up of all the Dark Knight Rises news doing the rounds »
Set vids, extras call, Cat suit…
Don't click this story if you're trying to avoid Dark Knight Rises Spoilers »
Bat bit leaks
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Modine to play Nixon in Batman movie »
"Full Metal Jacket" star is crooked politican
Batman coming to New Jersey? »
Batman coming to New Jersey?
www.superherohype.com reported a story that Christopher Nolan's "The Dark Knight Rises" may be looking to do some filming Read more…
Lost and Millennium actors join Dark Knight Rises »
primetime stars take on Batman
Joker joins The Dark Knight Rises!? »
Trio of new characters added to sequel
Here's Tom Hardy as Bane in The Dark Knight Rises! »
Bat vs Bronson!
The Dark Knight Start-of-Production Announcement »
The Bat. The Cat. The Freeman
Set pics from The Dark Knight Rises Online! »
Filming in London this week
Set pics from The Dark Knight Rises; Liam Neeson reprising Ra's al Ghul? »
Kermit Likes
Quick News – Lana Lang contender, Rock as Charley Pride, Batman locale »
All-new look!
Warner Bros officially names Cotillard and Gordon Levitt part of Dark Knight Rises »
Who're they playing?
Various Casting Bits : Dark Knight Rises, Odd Thomas, Wallflower, Big Sur, Liars All »
Oh, and Anna Faris is back.
Rescue Me's Franco gets a transfer to Gotham City »
Sunjata & more aboard "Dark Knight Rises"
Quick News – Life of Pi, Fright Night, Dark Knight Rises, Jeff Buckley Biopic »
Come check out my bits!
Why Dark Knight Rises is shooting in Pittsburgh »
Felines flock to City
Treat Williams offered roles in Dark Knight Rises, Tarantino Western »
The "Everwood" favourite making a comeback
Gary Oldman talks Dark Knight Rises »
Shooting dates, story, possible sequel
Gordon-Levitt as Falcone in Dark Knight Rises report false? »
Has Variety got it wrong?


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In Other News
Torrington girls soccer caps off season with annual banquet
Brady, Patriots roll over Lions for seventh straight win
College Sports
College football: Assessing threats for contenders in rivalry week
Litchfield Hall of Fame to induct 10 new members
College Sports
Top four unchanged in latest AP college football poll
West Virginia upsets UConn in Puerto Rico Tip-Off title game
SAN JUAN, PUERTO RICO >> Prior to Sunday night, the last time UConn faced an aggressive, choking, full-court pressure defense was nearly eight months ago against Louisville in the American Athletic Conference championship game. ...
Four UConn men's basketball players suspended
Omar Calhoun, Rakim Lubin, Dan Guest and Marcel Lewis were suspended for Sunday night's game due to a violation of team rules. Their status will be reevaluated when the Huskies return home. ...
College sports
A happy return to state for Cincinnati's Jake Golic
EAST HARTFORD >> The ball was tightly wrapped in Jake Golic's huge hands, and it was going to take an act of congress for the Cincinnati tight end to relinquish his grip.
College sports
UConn field hockey team wins national championship
For the second-straight season the UConn women's field hockey team finished as the nation's top team, defeating Syracuse 1-0 in the NCAA national championship game Sunday at College Park, Maryland.
Sunday Gravy: Ed McCarthy, West Haven power struggle reminiscent of Calhoun, UConn
This week's Sunday Gravy coming to you from Boston. Interesting side note from the 131st edition of "The Game." It's easier to be accepted into Harvard than it is to find a parking space within a two-mile radius of campus.
UConn Football
From bad to worse, UConn shut out at home by Cincinnati
EAST HARTFORD >> The day started with news leaking out about the departure of a pair of players expected to play major roles on the 2014 UConn football team. It continued to go progressively downhill from there.
Chip Malafronte: Yale is hurting after loss to Harvard, but Bulldogs are back
BOSTON >> Perhaps someday, the pain will subside and Yale can reflect justly on an otherwise remarkable season with pride. But Saturday's sting may linger for a while.
UConn Men's Basketball
No hard feelings between UConn's Ryan Boatright, West Virginia's Bob Huggins
SAN JUAN, Puerto Rico >> At some point prior to Sunday night's game, Ryan Boatright and Bob Huggins will likely cross paths.
Undefeated Harvard beats Yale to win Ivy League championship
CAMBRIDGE, Mass. >> The 131st edition of The Game will be remembered by The Finish.
UConn Women's Basketball
UConn women likely to put Creighton over a barrel
STORRS >> Creighton comes into Gampel Pavilion armed with one of the nation's best shooters, and with every intention of making a run at an NCAA tournament berth.
UConn Women's Basketball
Women's basketball preview capsule: No. 1 UConn vs. Creighton
The top-ranked UConn women's basketball team will take on Creighton on Sunday at Gampel Pavilion.
UConn Men's Basketball
Men's basketball preview capsule: No. 17 UConn vs. West Virginia
Seventeenth-ranked UConn will take on West Virginia in the championship game of the Puerto Rico Tip-Off tournament on Sunday in San Juan.
- 4 Litchfield County men indicted on federal fraud charges
- Bristol family adopts child in open Torrington ceremony
- Red Cross aids four Torrington families displaced by fire
- Adult, child hospitalized in Torrington as crews battle 2-alarm house fire
- Reibman's Torrington children's store closing after 60 years
- Study suggests genetic link for male homosexuality
- Benefit for Winsted native battling cancer to be held Nov. 30
- 4 things great marriages have in common


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- Northern Illinois to get $8.3 million for state construction projects
- Tree-lighting festival kicks off holiday season in Machesney Park
- Roscoe Boy Scout Troop's tree stand at new location
- Tips for selecting safe toys for kids this holiday season
- Prayer service for World AIDS Day Nov. 30
- Food Bank joins national #GivingTuesday movement
- Lee Hamilton: What lies ahead for Congress
- Rockford Public Schools faces $8.8 deficit, board OKs flat tax, HR chief
- Literary Hook: A holiday tradition: 'This Thanksgiving, Remember'
- Cold snap does not negate global warming
Literary Hook: In Print hosts book fair Sept. 10 at Barnes and Noble
By Christine Swanberg
Author and Poet
The In Print Professional Writers Group will offer "A World of Words: In Print" from 1 to 7 p.m., Saturday, Sept. 10, at Barnes and Noble, CherryVale Mall, 7200 Harrison, Rockford.
Featured book fair events include guest authors, book signings, readings, demonstrations and children's events.
Go to InPrintWriters.com for more detailed information.
Especially for poetry folks, here is the poetry schedule:
1:30-2:30 p.m. — Susanna Lang book signing
2:30-3:30 p.m. — Poetry readings
3:30-4:30 p.m. — Poetry discussion
4:30-5:30 p.m. — Christine Swanberg book signing
In Print is a professional writers' organization promoting creativity, commerce, craft and the community of writers (inprintwriters.com) and is an affiliate of the Chicago Writers Association (chicagowrites.org/).
In Print meets on the second Saturday of every month from 1 to 4 p.m. at the Cherry Valley Public Library (755 E. State St., Cherry Valley, 815-332-5161). Meetings include keynote speakers, workshops, critique sessions, writing prompts, editing services, and networking for published and unpublished writers of all genres. Members receive a calendar of conferences, contests and classes; a referral list of agents, and a monthly newsletter.
Yearly dues are $30. Guests may attend two meetings before joining. Contact email@example.com for more details.
Hope to see you at the book fair!
Christine Swanberg has published about 300 poems in 70 journals and anthologies. Her books include Who Walks Among the Trees with Charity (Wind Publishing, Kentucky), The Red Lacquer Room (Chiron Publishing, Kansas), The Tenderness of Memory (Plainview Press, Texas), Slow Miracle (Lakeshore Publishing, Illinois), Invisible String (Erie St., Illinois), Bread Upon the Waters (Windfall, Wisconsin) and Tonight on this Late Road (Erie St., Illinois).
From the Sept. 7-13, 2011, issue


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Name That Christmas Movie – Win a New Christmas Tree and Living Room Set
You could win a new Christmas tree and living room to enjoy it in.
You could win a new Christmas tree and living room to enjoy it in.
The latest shot in the battle to make people accept and love their bodies has been fired by a doll that could very well turn the toy industry on its ear.
Many local opportunities to see festive parades and get that Santa sighting!
Owensboro, where Kentucky starts Christmas!
Owensboro - Daviess County Christmas Parade - November 22, 2014 at 4:30 p.m. This parade w
The Bitterman Shoppes will be hosting a Christmas Open House on Friday and Saturday November 21st & 22nd. Kat stopped in this weekend and snapped some photos of what you can expect to find!
Get your pets' photo taken with Santa and help benefit the dogs of It Takes a Village Canine Rescue.
The holidays are right around the corner and it won't be long until decorations will be up and the festive holiday plants will be out on display. But what if you mistletoe has other capabilities other than making couples smooch?
The world-famous Budweiser Clydesdales, the symbol of quality and tradition for Anheuser-Busch since 1933, are scheduled to make several appearances in the area November 18 -23, 2014 including one on North Main Street in Evansville in conjunction with the Evansville's Christmas on North Main Parade. -Gail Knight Williams
In this age of superstores it is easy to forget to look at the treasure trove of local shops. Here are a few Owensboro stores and some of the unique Kentucky and Owensboro items available for the perfect gift
A certain ex-boyfriend of mine (who shall remain nameless but was an excellent gift giver) presented me with one of my most treasured possessions on Christmas of 2003. It was The Peanuts Holiday Special Box Set. If you know me at all, you know that Snoopy is my all-time favorite character.
My little sister had her first child this past Thursday, July 10th. We all held our phones up and took joyous pictures of each of us snuggling the little sweet fresh precious life and cried tears of joy together that we would have a new dear baby to love. I was especially happy because this one was not mine.
OK, so I know Christmas was a few weeks ago now, but people keep asking me how my Christmas was and what I got this year. Well, to answer the first question, my Christmas was great this year! And to answer the second, I got a LOT of toys and video games for Christmas! Here's a video I made highlighting some of the fun things Santa brought me this Christmas!
As if they weren't enough of a power couple already, now we find out they are even funny together.


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Getting Pregnant
Labor and Delivery
Baby and Beyond
Good grief. Can't use 'BMI', can't give a 'F' grade. How many more letters can we take out of the vocabulary of the school-aged?
I see obesity as a much larger epidemic than anorexia. They don't want to use the BMI figure for fear of creating an eating disorder??? Well look around you - Schools are filled with kids with eating problems. Just their problem is eating too much and/or eating the wrong stuff, with zero exercise.
If I had to choose one word, I'd go with harmful over helpful. This is something I expect from my child's doctor, not her school. Her doctor knows her, knows her historic growth cycles, and can offer real information about her health and nutrition instead of just "here's your number."
I'd rather see the money & energy being spent on BMI measurement go into an effort to get the kids to actually eat their more-nutritious school lunches rather than having kids go hungry all day & then pick up junk food after school. The grownups say, we need school lunches to be more nutritious, so the government is making it happen, and now the kids are throwing the fresh fruit & whole grain pasta into the trash. I saw this article the other day but couldn't post because DH was hogging the computer.
Some School Districts Quit Healthier Lunch Program - ABC News
The number of U.S. states in which a person can marry the person they love regardless of gender: 30 and counting!
Wow! The comments are vicious!
When I was in school we got weighed twice a year and the weight was written on our report cards. AFAIK, this never created an eating disorder in my peers or me. They screened for several things that they still screen for - scoliosis, hearing, vision. They also test cognitive ability with standardized, GATE, and special ed. testing.So Right! We got the letter in the mail too. Teachers should get weighed in too!Call me when they start getting rid of the fat teachers that set such a bad example for the students.
I just can't see it as a bad thing. It's data. It's educating parents as to what is a healthy weight and what is not. Every time I take my kids to the doctor, he or she tells us "you are in the ___ percentile on height; the ___ percentile on weight." Just like not every parent can/will take their child to the eye doctor, they don't all get to the doctor unless there is a problem. I would not take offense to a letter suggesting I see a physician re: my child's height/weight/vision/hearing/spine.
I think it is more harmful than helpful. I agree with the lady on the video that says that someone that has an obese child, knows that their child is obese. Just telling a child that they are "bad" is not going to make them better. The only thing I can see this accomplishing is more bullying and more eating disorders.
Stacy - The school that DH worked in last year had a program like that. They had all kinds of fresh fruits and veggies. Kids threw the food out in droves. They ended up setting up a table for the kids to put the unwanted food on for others to take.
The number of U.S. states in which a person can marry the person they love regardless of gender: 30 and counting!
Why is weight still the elephant in the room? There is this social stigma that is really silly when you think about it. You're not a healthy weight but we can't point out that you're not at a healthy weight because kids who didn't used to know you are fat will now know.
On a personal note, would your children's classmates care? Around here chunky seems to be the norm. But no one seems to notice or care.
Hmmm... I would not like to get a fat letter, actually I might like it since Robbie is so underweight...
Robbie's school is an APPLE school, we have to sign a family commitment to healthy eating and healthy lifestyles. When there are class parties there are healthy treats brought in, we do a candy exchange at Halloween (they get to pick a cheap toy), when we have hot lunches (2 times a year) they are healthy with all 4 food groups covered. They do random exercises during the day (do 10 jumping jacks and then get back to work as an example). The test the grade 5 class every year (when they take the weight, the kids do not see anything, it is hooked up to see in another room and they do a survey. The information is then averaged. Before the program started 3 years ago, 0% of the kids were getting 5 servings of fruits/vegetables a day, 5% were getting 1 hour of exercise/day (they have 30 minutes of phys ed daily), 60% spent 2+ hours on computer/tv/video games. In the past 3 years there has been a steady improvement.
Margaret (44)
Sean (38 )
Robbie (8 )
Bailey (April 2, 2011)
"The soul always knows what to do to heal itself. The challenge is to silence the mind." Caroline Myss
Obese And Overweight Teenagers More Likely To Have Considered Suicide
Teens Who Think They're Overweight More Likely To Try Suicide
Body Image Issues And Teen Suicide - CBS News
It would be far more beneficial to work time into the day for physical exercise and education on nutrition then to make kids who already feel really bad about themselves feel worse.


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No. 9 Men's Soccer Downs Villanova, 3-0
Musketeers' senior class becomes winningest in school history
BIG EAST Championship Match at PPL Park
The Musketeers fell 2-1 in the finale of the BIG EAST Tournament on Sunday, Nov. 16.
As A Sophomore (2013 - McKendree)
- Second Team All-GLVC
- Academic All-GLVC
- Game-by-Game Stats
Earned his second consecutive second team All-GLVC honor ... earned Academic All-GLVC Honors ... played in 16 games, making 15 starts ... led the team with seven goals, two assists and 16 points.
As A Freshman (2012 - McKendree)
- Second Team All-GLVC
- Game-by-Game Stats
Was a second team All-GLVC honoree ... played in 16 games, starting in 14 ... led the team with eight goals and was second on the team with five assists and 21 points ... scored his first collegiate goal and also tallied his first collegiate assist in a 4-1 win over Madonna on Aug. 30 ... recorded his first collegiate hat-trick with three goals in a 5-1 win at Kentucky Wesleyan on Oct. 21.
High School
Member of the New Zealand U17 World Cup qualifying team ... won the Golden Boot award as part of the the Mainland Federation team ... helped lead Nayland College to a runner-up finish in the Secondary School National Tournament.
Born January 14, 1994 ... son of Michael Ridsdale and Marie Calderbank ... comes form same hometown as incoming freshman Cory Brown ... also contacted by St John's.


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Hello and Scottoiler tip.
1 Attachment(s)
Hi, Im new to the group although I have been Lurking for a while. I have recently bought an xt 600 2003, and have read through many of the previous topics with interest. My biggest bugbear with the bike is the rusty downpipes, I m interested in replacing the whole system with stainless although I dont like the look of the price.
I have fitted a Scottoiler which apparently increases the life of the chain and a few colleagues have been raving about them. On fitting I must admit that I was quite impressed with the concept of only oiling the chain when the bikes in motion. The final delivery pipe was a big dissapointment, I found it difficult to position it precisely over the links, it seemed to have a mind of its own. After much headscratching, I decided to make the final delivery pipe from a spare length of copper brake line attached to the lower chaingaurd with a couple of homemade copper P-clips. If anyone else is having problems with final delivery give the brake line a try.
Jimmy B
I have a Hornet with a Scotoiler and I agree that the system is excellent for keeping the chain lubed up.
Be careful if you change the downpipes. I have a 2002 xt600 and the exhaust started blowing so I set about replacing with the stainless Motad header pipes. The problem is the 6mm studs that hold the pipes to the block. They are famous for causing problems by snapping on removal. 2 of my 4 sheared. One of the snapped off studs had to be drilled out and I ended up having to re-tap the thread to 8mm which is not unusual. A full weekend later I had the new pipes on!...Lots of swearing along the way. I hope the stainless will now outlast the bike but I will free them off now and then to be safe. The other problem was that the shoulders on the pipes remained pressed into the engine and limited access made subsequent removal difficult.
Best of luck...
Scottoiler delivery option
Hi,I'm also new to the site which is a real goldmine of info.
I fitted a Scottoiler Slipper block which was new out this year I think, it fits around the chain keeps it clean and lubed and works really well.
You might have to watch out it does'nt get dislodged if off road in the really
rough stuff though.
Oh Sun whereart thou!
|All times are GMT +1. The time now is 08:01.


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João Manuel Vieira Pinto (born 19 August 1971) is a Portuguese retired footballer who played mostly as a forward.
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As potential return of Cowboys' Josh Brent nears, mother of Jerry Brown pushes to bolster son's legacy
As potential return of Cowboys' Josh Brent nears, mother of Jerry Brown pushes to bolster son's legacy
Manny Pacquiao workout session - Behind the scenesView gallery
Manny Pacquiao workout session - Behind the scenes
Why is Manny Pacquiao fighting in Macau? Because of this rising star ...
Why is Manny Pacquiao fighting in Macau? Because of this rising star ...
Two-Minute Minor - The NHL's next monster contract goes to...View video
Two-Minute Minor - The NHL's next monster contract goes to...
The five worst teams in NFL historyView video
The five worst teams in NFL history
Manny Pacquiao spreads word about Christian-based project
Manny Pacquiao spreads word about Christian-based project
Matt Sandusky now an advocate for abuse preventionView video
Matt Sandusky now an advocate for abuse prevention
Yasmany Tomas' power has baseball in a frenzy
Yasmany Tomas' power has baseball in a frenzy
9 NFL games affected by weather, disasters or tragediesView gallery
9 NFL games affected by weather, disasters or tragedies
Three Periods: Maple Leafs need new approach more than new coach
Three Periods: Maple Leafs need new approach more than new coach
Oakland Raiders' crazy celebration saved by timely timeout
You'll have to excuse the Oakland Raiders for losing their minds celebrating late in Thursday night's 24-20 win over the Kansas City Chiefs.Shutdown CornerThu, Nov 20Kansas City20 - 24OaklandGame Recap
- AdChoicesStudent Debt ForgivenessSponsored
Student Loan Forgiveness: (800) 304-3124
Half the students who qualify for Student Loan Forgiveness never take advantage of the program. Call to see if you qualify: (800) 304-3124
Adrian Peterson says he won't use switch again, ponders new start
Vikings running back Adrian Peterson says he won't use a switch again and thinks he might need a fresh start elsewhere.Shutdown CornerSun, Nov 2310:00 AM PSTGreen Bay at MinnesotaPreview Game
Shuffle Up: Running back and tight end values, rest of season
Time to put the running backs and tight ends under the pricing gun, starting fresh.Roto Arcade
Winless Sixers force Papa John's to change pizza promotion
A Sixers "win" is now much less impressive than an actual win.Ball Don't LieFri, Nov 214:00 PM PSTPhoenix at PhiladelphiaPreview Game
Nike nixes tribute shoe plan after Iverson protest
Nike is pulling back on plans for a shoe that sneakily honors Allen Iverson after the former NBA superstar questioned whether the company could use his old number and team colors while he's endorsed by ...Associated Press
Pacquiao in need of signature win against Algieri
MACAU (AP) — Boxing is a hurt business, as Manny Pacquiao found out when he ended up face down on the canvas after being knocked out just three fights ago by Juan Manuel Marquez.Associated Press27 mins ago
Dodgers get reliever Joel Peralta in four-player trade with Rays
The Dodgers hope Peralta, 38, can bring a bit of stability to their bullpen.Big League Stew
- AdChoicesInstant CheckmateSponsored
Forget 'Googling' Their Name, Use This New Site!
A new site has made it easy to find out the truth about the people you already know and people you meet! Simply type in a name and select a state!
Mario Williams plows Jim Kelly's driveway during Buffalo snowpocalypse
Jim Kelly, like the rest of Buffalo, was snowed in during an incredible blizzard pelting the city. Mario Williams showed up to help.Shutdown Corner
The unintentionally hilarious Claude Giroux injury drama timeline
Here's a look back at the odd journey the Flyers captain took from significant injury to starting lineup, made even odder by the media frenzy and social media inanity that surrounded it.Puck Daddy
Toughest fantasy decisions of Week 12
Rotoworld: Adam Levitan joins Jenna Corrado to break down the toughest fantasy calls heading into Week 12 of the NFL season, including if Tre Mason is a reliable start against the Chargers.NBC Sports
'The rebuttal' provides rare glimpse inside Tiger Woods' mind
Tiger Woods' response to Dan Jenkins satire was a fascinating look at the real, unvarnished megastar.Yahoo Sports
Kansas State's win against WVU was the best thing for the Big 12
The conference knows that it will have at least two teams positioned to make a run at the playoff in the final two weeks and potentially three.Dr. Saturday
Keith Olbermann says Giancarlo Stanton's contract is a scam
Are the Marlins trying to trick us all again with Stanton's $325 million deal? Olbermann thinks so.Big League Stew
- AdChoicesLendingTreeSponsored
An Extremely Brilliant Way To Pay Off Mortgage
If you own a home and pay for a mortgage, you could reduce your payments by an average of $4,100 a year. Here is how it works.
Vince Carter breaks down in tears after watching Toronto's tribute to him (Video)
The former Raptor had an up and down time in Toronto.Ball Don't LieFri, Nov 214:30 PM PSTMilwaukee at TorontoPreview Game
See 'Sin City Saints' Season 1 Photos (8 photos)
From executive producer Mike Tollin (One Tree Hill, Smallville, and Varsity Blues) and two-time Emmy nominated director Bryan Gordon (Curb Your Enthusiasm, The Office, Party Down), Sin City Saints is an off-beat comedy set in the front office of a fictional pro basketball expansion team. The show…Yahoo TV
Phil Mickelson Has Shed A Ton Of Weight On The LeBron Diet
After a rough 12 months in which he fell from...Business Insider
Pirates designate Ike Davis for assignment
PITTSBURGH (AP) -- The Pirates have designated first baseman Ike Davis for assignment and have added pitcher Jameson Taillon to the 40-man roster.The Associated Press
NFL against the spread picks: The Chiefs' really bad spot
We should be able to point to some good reason why an 0-10 team beats a 7-3 team that has emerged as one of the safest picks every week in the NFL.Shutdown Corner
Slava Voynov charged with felony for 'corporal injury to spouse with great bodily injury'
The Los Angeles Kings defenseman was charged in regards to a domestic violence case involving his wife.Puck Daddy
- AdChoicesThe Easy Loan SiteSponsored
Homeowners are in for an Unexpected Surprise
An often overlooked method to paying off your mortgage will have banks across the country in a panic. You'll want to see this if you're a homeowner.
Denver's linemen break silence, vow to get better
ENGLEWOOD, Colo. (AP) -- Orlando Franklin ambled into the locker room Thursday and loudly announced, ''I'm available to talk about the situation.''The Associated PressSun, Nov 231:25 PM PSTMiami at DenverPreview Game


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November 26, 2014
KANSAS CITY, Mo. – The Kansas City Chiefs announced on Tuesday that the club has signed defensive tackle Kyle Love and released defensive tackle Anthony Toribio.
Love (6-1, 315) has played in 41 games (25 starts) in three NFL seasons with the New England Patriots (2010-12). He has tallied 65 tackles (36 solo), 5.5 sacks (-22.0 yards), one fumble recovery and one pass defensed. Love was acquired via waiver claim by the Jacksonville Jaguars on May 16, 2013 and released by Jacksonville prior to the start of the regular season. He originally entered the NFL as an undrafted free agent of the Patriots in 2010. Love played in 44 games at Mississippi State, finishing his career with 71 tackles. Born in the Republic of South Korea, the Fairburn, Ga., native prepped at North Clayton High School in College Park, Ga.
Toribio (6-1, 315) has seen action in 25 contests in five NFL seasons with the Green Bay Packers (2009) and Kansas City Chiefs (2010-13). He owns 12 career tackles (nine solo). He originally joined Kansas City in 2010 off a waiver claim from the Green Bay Packers. Toribio spent the 2011 season on the Chiefs practice squad roster. He appeared in one game for Green Bay in 2009 after originally entering the NFL as a rookie free agent with Miami in 2008. The Miami, Fla., native was a four-year letterman at Carson-Newman.
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OK, we're officially in a recession. So what better time for Grazia's Easy Chic Week. Killer cocktail dresses at Coast, hot heels at Faith, the essential little lace dress at Oasis, winter cashmere at Uniqlo, tux dressing at Principles, A/W essentials at Wallis and designer goodies galore including these Marc by Marc Jacobs boots at Flannels - all with a massive 20% discount. Quick - get thee to your local newsagent to pick up your latest Grazia, like now. And enjoy your (kinda) guilt free shopping!
Get a 20% discount off Oasis, Coast, Uniqlo and more...
21 October 2008
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Today, House Appropriation's overflow room was overflowing. Concerned citizens were lining up, some in wheelchairs, to testify about the pain that the proposed budget cuts would inflict. In January, when Chairman Jim Pitts laid out his draconian budget proposal, he called it "a starting point." But as the days wear on, it looks less and less likely that major improvements can be made. While some members of the Senate might be willing to look at an increase in registration fees and in the gas tax, House Speaker Joe Straus is telling people that "under no circumstances" will a tax bill be considered. And why would he advocate one? As one senator reminded me today, why should lawmakers "fall on their swords" to support a tax bill that Gov. Rick Perry would veto, with glee?
Soon, and then too often, we will hear lawmakers resorting to that well-worn phrase signaling desperation: "You can't put lipstick on a pig." It's going to be that kind of budget.
While the House will pass the budget bill first, Lt. Gov. David Dewhurst has told confidants that he has a fall-back budget plan that, while not pretty, spreads the pain around as evenly as possible. The concept is simple: "vital" services keep their current levels of funding, with no adjustments for inflation, patient loads or school enrollment. It has a "Hail Mary" aspect to it, postponing most cuts until 2013, in hopes that the state's economic growth will rebound and produce more revenue for the state treasury that year.
Also today, Dewhurst laid out his legislative plan, which will be carried by Sen. Jane Nelson, to encourage change in Texas medical care delivery by protecting insurance companies and doctors' and hospital groups from anti-trust charges. Proponents believe the legislation will save the state money by allowing the creation of new medical consortiums that can deliver better outcomes for less money. But can the state count on savings in this biennium? It takes a while to get these reforms going and see their effects. Republicans in Washington are complaining a lot about phantom savings in "Obamacare;" don't be surprised if Republicans in Texas use "phantom savings" to help balance the state's budget.
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Medical examiner investigating infant death
The medical examiner says indications are the baby was sleeping with his mother and a sibling.
The medical examiner says indications are the baby was sleeping with his mother and a sibling.
The FCC is nearing a decision on how far to go to protect Internet consumers from deals between broadband providers and content companies.
The post office in De Pere is named the Cpl. Justin D. Ross Post Office.
The FBI said it was leading a multi-agency investigation into the breach, which took effect in mid-September.
Forty-two-year-old Salah Salahadyn pleaded guilty to robbery in October.
Hundreds of people gathered for the memorial Sunday night for Laylah Petersen, who was killed Thursday.
The DNR says the software can help engineers, nutrient management planners, county conservationists, researchers and farmers address polluti…
Perfect Holiday snack recipe.
We're at Salon Aura for our Monday Morning makeover! Check out how our friends from the salon transformed Jeannie's look!
Segment sponsored by The Bulk Priced Food Shoppe.


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Category: Air & Climate ×
Subcategory: Air Monitoring ×
Keyword: ldar ×
Employees number: 11-100 ×
Location: USA ×
Location: Nevada ×
Hetek Solutions Inc.
based in London, ONTARIO (CANADA)
At Hetek, we consider the environment our most important resource. That`s why we deliver efficient, safe, and environmentally sound services that reduce waste and maximize your water and energy assets. And because conservation begins with each one of us, Hetek also provides hands-on, personal training in all areas of safety and resource management.Hetek Solutions Inc. ...
Gratton Arabia Ltd.
based in Mississauga, ONTARIO (CANADA)
Gratton Arabia Ltd. offers specialized and niche services that address, energy management, employee safety, training and corporate compliance. Gratton Arabia Ltd. is an electrical safety oriented company, joining together various aspects of the electrical industry, such as engineering, analysis, design, installation, field services, standards, manufacturing and supply, ...
based in Chesterfield, MICHIGAN (USA)
Argus-Hazco offers an extensive line of industrial health, safety and environmental monitoring equipment, supplies and services. North American Coverage supported by our network of local service centers ensures that your job has the equipment and supplies you need—when and where you need them. Our diverse and comprehensive rental fleet provides you with the flexibility to ...
based in Birkenhead, UNITED KINGDOM
a1-cbiss are an ISO accredited, environmental monitoring systems supplier with expertise in the design, manufacture and integration of fixed and portable environmental monitoring solutions: MCERTS accredited Continuous Emissions Monitoring Systems; Gas Detection & Analysis Instrumentation; and Sequential Sampling Systems. a1-cbiss offer complete turnkey solutions ...
DOD Technologies Inc
based in Crystal lake, ILLINOIS (USA)
DOD Technologies is a leading global supplier of integrated gas detection systems and solutions. Our innovative products, services and systems protect, support and save lives. Semiconductor, Solar, Specialty Chemical, Polyurethane foam, Wastewater, Engineered wood and emergency services industries have trusted in DOD Technologies since our inception. With over 100 years ...
GOW-MAC Instrument Co.
based in Bethlehem, PENNSYLVANIA (USA)
GOW-MAC Instrument Co. is a leading manufacturer of gas chromatographs, gas analyzer packages and specialty gas handling equipment for user-specified applications for industrial gas, laboratory, and educational applications. Our line of instruments includes gas chromatographs and detectors (TCD, DID, FID, PID, FPD), gas analyzers (trace, binary and toxic), gas leak ...
US Environmental Rental Corporation
based in Waltham, MASSACHUSETTS (USA)
We specialize in environmental equipment rentals, sales and service. Competitive pricing in all categories from air, soil, water and survey and then more. We will provide top of the line equipment with the best service in the industry. We take pride in our work and will go the extra step to help out however we can. Give us a try an experience the friendly service that we ...
Monitor Technologies LLC
based in Elburn, ILLINOIS (USA)
Monitor Technologies LLC is a dependable manufacturer of material level measurement and monitoring instruments, solids flow monitors, particle emission monitors and bin aeration devices for the worldwide powder and bulk solids market. The experience and market knowledge gained from over fifty years in business has earned Monitor a reputation for offering high quality, ...
Enviro Technology Services plc
based in Stroud, UNITED KINGDOM
Enviro Technology Services plc (ET) was founded in 1983. Now led by MD, Mike Webley the company is active in 45 countries with a multi-million pound turnover, ET has evolved over 31 years to become a leading provider of air quality and emissions monitoring systems and servicing across the globe.
Apollo Safety Inc.
based in Fall River, MASSACHUSETTS (USA)
Apollo Safety has been serving the safety industry since John and Tracy Carvalho started the business in February 1995. Apollo has grown to be the leading safety and technical services equipment provider in New England and serving by courier across the Untied States and Internationally. Apollo Safety understands that safety is much more than just a commodity. Our focus ...
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So I have say two projects, project A and project B. Project A contains a main class which I want to run. Project B contains some classpath resources which are required and also has a reference to Project A. If I run the main class in Project A directly, it will only run it in the context of that project and won't include the classpath resources as they're in Project B.
But if I try to run Project B, I can't select the main class (as it doesn't exist in that project) and if I try right-clicking the project -> Run As -> Java Application, it doesn't come up with that main class as a valid main class. (It does come up in the similar dialog for Project A.)
This seems to be a common problem from Eclipse; I found this related question but this person is not using Maven. Also, they accepted an answer which referenced project B from project A, which breaks the encapsulation I am trying to achieve here: Eclipse 3,6 (Helios) referenced project has main method, cannot access from run config when targeting referencing project