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The bone morphogenetic protein (BMP) signaling pathway regulates multiple developmental and homeostatic processes. Mutations in the pathway can cause a variety of somatic and hereditary disorders in humans. Multiple levels of regulation, including extracellular regulation, ensure proper spatiotemporal control of BMP signaling in the right cellular context. We have identified a modulator of the BMP-like Sma/Mab pathway in C. elegans called DRAG-1. DRAG-1 is the sole member of the repulsive guidance molecule (RGM) family of proteins in C. elegans, and is crucial in regulating body size and mesoderm development. Using a combination of molecular genetic and biochemical analyses, we demonstrate that DRAG-1 is a membrane-associated protein that functions at the ligand-receptor level to modulate the Sma/Mab pathway in a cell-type-specific manner. We further show that DRAG-1 positively modulates this BMP-like pathway by using a novel Sma/Mab-responsive reporter. Our work provides a direct link between RGM proteins and BMP signaling in vivo and a simple and genetically tractable system for mechanistic studies of RGM protein regulation of BMP pathways.
Bone morphogenetic proteins (BMPs) belong to the transforming growth factor β (TGFβ) superfamily of ligands and the BMP signaling pathway plays roles in multiple developmental and homeostatic processes (Wu and Hill, 2009). Malfunction of the pathway causes many somatic and hereditary disorders in humans, including cardiovascular diseases and cancer (Gordon and Blobe, 2008; Massagué, 2008). All BMP family members share a common mode of signal transduction (Shi and Massagué, 2003). Upon binding of the BMP ligands, the type II receptor phosphorylates the type I receptor, which then phosphorylates the receptor-regulated SMADs (R-SMADs). These activated R-SMADs then complex with common-mediator SMADs (co-SMADs) and enter the nucleus where the complex participates in regulation of downstream gene expression. Multiple levels of regulation, including extracellular regulation, ensure proper spatiotemporal control of BMP signaling in the right cellular context (Massagué and Chen, 2000; Balemans and Van Hul, 2002; Moustakas and Heldin, 2009; Umulis et al., 2009). In particular, a family of repulsive guidance molecules (RGMs) has recently been found to bind different BMPs with high affinity and act as BMP co-receptors in tissue culture experiments (Samad et al., 2005; Babitt et al., 2005; Babitt et al., 2006).
RGM family proteins are GPI (glycosylphosphatidylinositol)-linked membrane-associated proteins (Corradini et al., 2009; Severyn et al., 2009). Vertebrates have four RGMs: RGMa, RGMb (DRAGON), RGMc (hemojuvelin or HJV) and RGMd (Camus and Lambert, 2007). RGMa, RGMb and RGMc can bind selected BMP molecules as well as type I and type II BMP receptors to enhance BMP signaling in tissue cultures (Babitt et al., 2005; Babitt et al., 2006; Samad et al., 2004; Samad et al., 2005; Xia et al., 2008; Xia et al., 2010; Andriopoulos et al., 2009). Overexpression of mouse RGMb in Xenopus embryos can augment SMAD-1-induced mesodermal and endodermal marker expression (Samad et al., 2005). Several lines of in vivo evidence suggest that RGMc acts as the co-receptor for BMP6 in regulating iron metabolism in mice (Babitt et al., 2007; Xia et al., 2008; Andriopoulos et al., 2009). However, there is, as yet, no evidence showing that RGMa and RGMb are required for modulating BMP signaling in vivo.
In this study, we describe the identification and functional characterization of the sole C. elegans RGM homolog DRAG-1 in modulating a BMP-like signaling pathway. There are two TGFβ-related pathways in C. elegans, a TGFβ-like pathway that controls dauer formation and a BMP-like Sma/Mab pathway that regulates body size, male tail formation, mesoderm development and innate immunity (Savage-Dunn, 2005; Foehr et al., 2006; Moustakas and Heldin, 2009; Partridge et al., 2010). The ligand of the Sma/Mab pathway is a BMP-like molecule DBL-1 (Morita et al., 1999; Suzuki et al., 1999). Additional members of this pathway include the type I receptor SMA-6 (Krishna et al., 1999), type II receptor DAF-4 (Estevez et al., 1993), and SMADs SMA-2, SMA-3 and SMA-4 (Savage et al., 1996). Loss-of-function mutations in any of these pathway members cause small body size and male tail sensory ray formation defects.
The Sma/Mab pathway also plays a role in patterning the C. elegans postembryonic mesoderm. The hermaphrodite postembryonic mesodermal M lineage arises from a single pluripotent precursor cell, the M mesoblast. During larval development, the M mesoblast divides to produce a dorsal lineage that gives rise to macrophage-like coelomocytes (CCs) and striated bodywall muscles (BWMs), and a ventral lineage that produces BWMs and the sex muscle precursor cells, the sex myoblasts (SMs; Fig. 1C) (Sulston and Horvitz, 1977). We have previously shown that mutations in the schnurri homolog sma-9 lead to a dorsal-to-ventral fate transformation in the M lineage (Foehr et al., 2006; Foehr and Liu, 2008). Furthermore, mutations in the core components of the Sma/Mab pathway suppress the dorsoventral patterning defects of sma-9 mutants, suggesting that SMA-9 functions by antagonizing Sma/Mab signaling to pattern the postembryonic mesoderm along the dorsoventral axis.
Among suppressors of the sma-9 phenotype, we detected a mutation in a novel locus that we have named drag-1. drag-1 mutants exhibit only a subset of phenotypes seen in the Sma/Mab pathway mutants. We show that drag-1 encodes the sole RGM homolog in C. elegans. We also show that DRAG-1 is a membrane-localized protein that functions at the ligand-receptor level in the Sma/Mab pathway to regulate body size and mesoderm patterning. Using a novel Sma/Mab-responsive reporter, we demonstrate that DRAG-1 positively modulates Sma/Mab signaling. Our work establishes a direct link between RGM proteins and BMP signaling in vivo and provides a simple and genetically tractable system for mechanistic studies on RGM protein regulation of BMP pathways in vivo.
C. elegans strains
Strains were kept at normal conditions, as described by Brenner (Brenner, 1974). Analyses were performed at 20°C unless otherwise noted. The following mutations and integrated transgenes were used. Linkage group II (LGII): sma-6(jj6); LGIII: daf-7(m62), daf-7(e1372), daf-4(m72), sma-3(jj3), lon-1(e185), ccIs4438(intrinsic CC::gfp); LGIV: daf-1(m40), daf-1(m213); LGV: dbl-1(wk70); LGX: lon-2(e678), sma-9(cc604), sma-9(cc606), sma-9(wk55). Other strains used included VF1: unc119(ed3); gfEx[phmt-1p::gfp, unc-119(+)] (Schwartz et al., 2010); VF12.3: hmt-1(gk161) III; gfIs1[hmt-1::gfp, unc-119(+)].
Molecular analysis of drag-1
The molecular lesion of jj4 was identified by sequencing PCR fragments spanning the entire genomic region of Y71G12B.16. To determine the drag-1 cDNA sequence, the ProQuest cDNA library (Invitrogen) was used for PCR amplification using primers ForwLongpPC-86 (corresponding to vector sequences 5′-TATAACGCGTTTGGAATCACTACAGGGATGTTTAATACCAC-3′) and DS147 (complementary to sequences spanning the predicted stop codon of drag-1 5′-TCAGCATAACAATGATAAAAGAGCAAAAAAAAAG-3′). The PCR fragments were cloned into the pCR-Blunt II-TOPO vector (Invitrogen). Three of the ten resulting clones sequenced contained the entire Y71G12B.16 open reading frame (ORF) as well as additional 5′UTR sequences. One clone (pDS9.10) has 32 bp of 5′UTR sequences immediately upstream of the ATG (Y71G12B.16b; Genbank HM154524), whereas the other two clones (pDS9.5 and pDS9.6) contain 60 bp 5′UTR sequences (Y71G12B.16a; Genbank HM154523).
Plasmid constructs and transgenic lines
drag-1 reporter constructs
Four kilobases of the drag-1 upstream sequence (−3977 to −1), sequences from immediately downstream of the ATG until the end of the first intron of drag-1 (4 to 1123), the entire drag-1 protein-coding region and 1 kb of the drag-1 downstream sequences were PCR-amplified using N2 genomic DNA as a template. The PCR products were used to generate the following reporter constructs for analyzing the expression pattern of drag-1: pDS14, 4 kb drag-1p::4xNLS::gfp::unc-54 3′UTR; pDS15, 4 kb drag-1p::drag-1 1st intron::4×NLS::gfp::unc-54 3′UTR (transgenic animals carrying either pDS14 or pDS15 gave identical gfp expression patterns; data not shown); pDS27, 4 kb drag-1p::drag-1 genomic ORF::1 kb drag-1 3′UTR; pCXT15, 4 kb drag-1p::drag-1 1st intron::drag-1 cDNA::unc-54 3′UTR; pJKL849, 4 kb drag-1p::drag-1 genomic ORF::gfp::1 kb drag-1 3′UTR. GFP from the Fire Lab Vector pPD95.75 (http://www.addgene.org) was inserted between amino acid (aa) 395 and aa396 of the drag-1 coding region, just prior to the cleavage site of the putative C-terminal pro-peptide.
Constructs for tissue-specific expression of drag-1
pJKL928, myo-2p::drag-1 cDNA::unc-54 3′UTR; pJKL931, elt-2p::drag-1 cDNA::unc-54 3′UTR; pJKL933, elt-3p::drag-1 cDNA::unc-54 3′UTR; pJKL935, rol-6p::drag-1 cDNA::unc-54 3′UTR; pDS20, hlh-8p::drag-1 cDNA::unc-54 3′UTR; pCXT148, hlh-8p::drag-1 cDNA::1 kb drag-1 3′UTR.
Constructs for structure-function analysis of drag-1 (derived from pJKL849)
pCXT92 (DelC::GFP), drag-1::gfp with the putative C-terminal pro-peptide (aa387-408) deleted; pCXT115 (DelN::GFP), drag-1::gfp with the N-terminal signal peptide (aa2-23) deleted; pCXT116 (DelNC::GFP), drag-1::gfp with both the N-terminal signal peptide (aa2-23) and the C-terminal pro-peptide (aa387-408) sequences deleted; pCXT94 (LIN-12TM::GFP), drag-1::gfp with the C-terminal pro-peptide (aa387-408) replaced by the LIN-12 transmembrane domain (aa907-934 of LIN-12) (Yochem et al., 1988).
Other reporters
pJKL840, sma-6p::nls::rfp::lacZ::unc-54 3′UTR [containing 3 kb of the sma-6 promoter (−3094 to −1)].
Generating transgenic animals
Transgenic animals were generated using the plasmids pRF4 (Mello et al., 1991), pC1 (pha-1 rescuing construct) (Granato et al., 1994), pJKL449 (myo-2p::gfp::unc-54 3′UTR) (Jiang et al., 2009) or LiuFD61(mec-7p::mRFP) (Amin et al., 2009) as markers.
Reverse-transcription PCR (RT-PCR)
Total RNA was extracted from mixed-stage wild-type and drag-1(jj4) worms using an RNA extraction kit (Qiagen). Reverse transcription was performed using the Superscript III First Strand Synthesize Kit (Invitrogen) following the manufacturer's instructions. Primers used for amplifying the wild-type and jj4 cDNAs were CXT49 (annealing to exon 1, forward) and CXT50 (annealing to exon 3, reverse). The resulting PCR fragments were purified and sequenced using primer CXT67.
Body size measurement
Hermaphrodite animals at the gravid adult stage were collected and treated with hypochlorite. The resulting embryos were allowed to hatch in M9 buffer at 16°C. Synchronized L1s were plated onto nematode growth medium (NGM) plates and allowed to grow for 24, 48, 72 or 96 hours, respectively, before they were washed off the plates, treated with 0.3% sodium azide and mounted onto 2% agarose pads. Images of the worms were taken on a Leica DMRA2 compound microscope with a Hamamatsu Orca-ER camera using the Openlab software (Improvision). Body length of each animal was measured using Openlab software. Subsequent statistical analyses were performed using Microsoft Excel.
Dauer formation assays
Frequency of dauer formation was assessed under non-dauer-inducing conditions as described (Vowels and Thomas, 1992). Five to ten adult hermaphrodites were placed on a 6 cm NGM plate at the test temperature. After a short period of egg-lay (less than 12 hours at 25°C), the parents were removed from the plates and their progeny were allowed to develop at a given temperature. When the non-dauer worms became young adults, the numbers of dauers and non-dauers on each plate were counted.
Immunofluorescence staining
Animal fixation, immunostaining, microscopy and image analysis were performed as described previously (Amin et al., 2007). Guinea pig anti-FOZI-1 (1:200) and goat anti-GFP (Rockland Immunochemicals; 1:1000) were used. All secondary antibodies were from Jackson ImmunoResearch Laboratories and used at a dilution of 1:50 to 1:200.
Fractionation experiment
Transgenic worms used in the fractionation experiment were generated in the pha-1(e2123ts) mutant background and maintained at 25°C. Worms were harvested in M9 and resuspended in lysis buffer [50 mM Tris-HCI, pH 7.5; 10% glycerol; 10 mM 2-mercaptoethanol, 1 mM phenylmethylsulfonyl fluoride and 1 Complete Mini Protease Inhibitor Cocktail Tablet (Roche) in 50 ml lysis buffer] so that the ratio of worm volume to buffer volume was 1:1. Worms were sonicated until around 90% of the worms were lysed as assessed by microscopy. Debris was cleared by centrifugation at 4000 g for 10 minutes at 4°C. The supernatants were centrifuged at 115,000 g for 1 hour. The resulting supernatant was the soluble fraction. The pellet, which contains microsomal membrane proteins, was further cleared from soluble proteins by two more rounds of centrifugation (115,000 g) and resuspension in lysis buffer. The resulting pellet was reconstituted in the same volume of lysis buffer as the soluble fraction. Micro BCA Protein Assay Reagent Kit (Pierce) was used to measure the protein concentration. Thirty micrograms per lane of samples were loaded onto a 7% SDS-PAGE. GFP fusion proteins were detected by western blotting using goat anti-GFP antibodies (Rockland Immunochemicals; 1:1000).
Generating the RAD-SMAD and BAD-SMAD reporters
Oligonucleotides containing the wild-type (RAD-SMAD) or mutant (BAD-SMAD) SMAD binding sites and StyI restriction sites at each end were treated with T4 polynucleotide kinase, annealed and ligated. Concatenated oligonucleotides were cloned into the Fire Lab Vector L3135 (http://www.addgene.org). Upon confirmation of the plasmid sequence, their DNA was injected into N2 worms using mec-7p::mRFP as a co-injection marker. The RAD-SMAD transgene was then integrated into the genome via gamma-irradiation. Seven integrated lines were obtained, out-crossed and mapped to specific chromosomes. All lines showed similar GFP expression patterns. Two of them, LW2433: jjIs2433[pCXT51(RAD-SMAD) + LiuFD61(mec-7p::mRFP)] X and LW2436: jjIs2436[pCXT51(RAD-SMAD) + LiuFD61(mec-7p::mRFP)] (I or IV) were used for subsequent analysis. The BAD-SMAD transgenes were kept in high-efficiency extra chromosomal lines.
drag-1 mutants exhibit a subset of the phenotypes seen in mutants in the Sma/Mab pathway
In a mutagenesis screen for suppressers of the sma-9(cc604) M lineage phenotype (Foehr et al., 2006), we isolated the Sma/Mab pathway mutants as well as a single-locus recessive mutation, jj4. Unlike sma-9(cc604) animals, which lack both M lineage-derived coelomocytes (CCs; Fig. 1A,D), 98.5% (n=201) of jj4; sma-9(cc604) animals had both M-derived CCs (Fig. 1B,C). jj4 also suppressed the M lineage phenotypes of two other sma-9 alleles, cc606 and wk55 (data not shown), suggesting that this suppression is not allele-specific. Like the Sma/Mab pathway mutants, jj4 mutants by themselves did not have any M lineage defect when separated from the cc604 mutation (Fig. 1C). However, jj4 mutants were smaller than wild-type N2 worms throughout larval development, although they were not as small as the dbl-1(wk70) null mutants (Fig. 1E-G). Unlike the Sma/Mab pathway mutants, jj4 mutant males could mate and did not exhibit any male tail patterning defects (Fig. 1H,I). Thus, jj4 mutants exhibit some, but not all, phenotypes of the Sma/Mab pathway mutants.
We mapped the jj4 mutation to the left arm of chromosome I, a region not previously known to contain any genes involved in body size regulation. Thus jj4 represents a novel locus required to regulate body size. We named the locus drag-1 (see below).
drag-1 functions in the Sma/Mab pathway, possibly at the ligand-receptor level, to regulate body size
Because drag-1(jj4) mutants and the Sma/Mab pathway mutants share similar phenotypes in body size and mesoderm patterning, we tested whether drag-1 functions in the Sma/Mab pathway. We generated double mutants between jj4 (a putative null, see below) and mutations in various Sma/Mab pathway components (Fig. 2A) and measured their body sizes.
As shown in Fig. 2B, double mutants between jj4 and the sma-3(jj3) null mutation were as small as sma-3(jj3) single mutants. Similarly, jj4; sma-6(jj1) and jj4; dbl-1(wk70) double mutants were as small as sma-6(jj1) and dbl-1(wk70) single mutants (both are null mutants), respectively. These observations suggest that jj4 is probably compromised for Sma/Mab pathway function, rather than affecting a pathway that functions in parallel to the Sma/Mab pathway, in regulating body size.
To further characterize the role of drag-1 in the Sma/Mab pathway, we generated the following double mutants: jj4; lon-1(e185), jj4; lon-2(e678) and jj4; dbl-1(ctIs40). e185 is a strong loss-of-function mutation in lon-1, a known downstream target of the Sma/Mab pathway (Maduzia et al., 2002; Morita et al., 2002). e678 is a null mutation in lon-2, which encodes a member of the glypican family of heparan sulfate proteoglycans and acts as a negative regulator of the Sma/Mab pathway (Gumienny et al., 2007). dbl-1(ctIs40) is a strain that overexpresses dbl-1 (Suzuki et al., 1999). As shown in Fig. 2B, jj4; lon-1(e185) mutants were as long as lon-1(e185) mutants. However, jj4; lon-2(e678) and jj4; dbl-1(ctIs40) double mutants showed intermediate body size between jj4 single mutants and lon-2(e678) or dbl-1(ctIs40) single mutants. These results suggest that drag-1 is likely to act upstream of lon-1, but in parallel to lon-2 and dbl-1. Taken together, our genetic epistasis results are consistent with drag-1 functioning at the ligand-receptor level in the Sma/Mab pathway to regulate body size.
drag-1 interacts genetically with the dauer pathway
In addition to the Sma/Mab pathway, C. elegans has another TGFβ pathway that regulates dauer development (Savage-Dunn, 2005). drag-1(jj4) animals did not exhibit any constitutive or deficient dauer phenotypes on their own (Table 1). Animals carrying a deletion allele of drag-1(tm3773) (see below), did not exhibit any dauer phenotypes either (Table 1). However, drag-1(jj4) enhanced the dauer-constitutive (Daf-c) phenotype of certain daf-7 and daf-1 alleles at the permissive temperature. daf-7 and daf-1 encode the ligand and the type I receptor of the dauer pathway, respectively (Georgi et al., 1990; Ren et al., 1996). As shown in Table 1, although drag-1(jj4) did not affect the Daf-c phenotype of daf-7(m62) animals at all temperatures tested, drag-1(jj4) moderately enhanced the Daf-c phenotype of daf-7(e1372) mutants at 16°C. Similarly, drag-1(jj4) moderately enhanced the Daf-c phenotype of daf-1(m213) mutants at 16°C, but significantly enhanced the Daf-c phenotype of daf-1(m40) mutants at both 16°C and 20°C. The allele-specific and moderate enhancement of Daf-c phenotypes of daf-7 and daf-1 mutants by drag-1(jj4) are consistent with low levels of crosstalk between the Sma/Mab and dauer pathways, as suggested by previous observations on the genetic interactions between sma-6 and daf-7 and daf-1 mutations (Krishna et al., 1999).
drag-1 encodes the C. elegans homolog of the RGM family proteins
drag-1(jj4) was mapped to the left arm of chromosome I between Y92H12A.3 and Y71G12B.24 via snip-SNP mapping (Wicks et al., 2001). RNAi injection for 15 likely candidates among the 24 predicted genes located in this region showed that Y71G12B.16(RNAi) led to a small body size phenotype and the suppression of the sma-9(cc604) M lineage phenotype. For simplicity, we will refer to the suppression of the sma-9(cc604) M lineage phenotype by jj4 as the SOSMLP phenotype of jj4 in the remainder of the manuscript. Several additional lines of evidence indicated that jj4 is a loss-of-function allele of Y71G12B.16. First, DNA fragments containing the genomic DNA sequence of Y71G12B.16 rescued both the small body size and the SOSMLP phenotype of jj4 mutants (Fig. 3C,D; Fig. 5O). Second, sequencing of Y71G12B.16 in jj4 mutants revealed a T-to-A single nucleotide mutation in the splicing donor site in the first intron (Fig. 3C). RT-PCR analysis (see Materials and methods) showed that this mutation led to the production of three aberrant drag-1 transcripts, all producing mutant peptides containing the first 16 amino acids of DRAG-1 followed by 9-49 additional random amino acids (data not shown). Third, a deletion allele of drag-1, tm3773, led to the same body size and SOSMLP phenotypes as jj4 mutants (Fig. 1G; Fig. 3C,D; Fig. 5O).
We isolated two cDNA variants for Y71G12B.16, Y71G12B.16a and Y71G12B.16b, which share the same coding sequences and differ only in the lengths of their 5′UTRs (see Materials and methods). The predicted Y71G12B.16 protein is 408 aa long. It contains a predicted N-terminal signal peptide, a partial von Willebrand factor D (vWF type D) domain, a hydrophobic region and a C-terminal region that meets the criteria of a pro-peptide that is cleaved off and replaced by a glycosylphosphatidylinositol (GPI) anchor during post-translational processing (Chatterjee and Mayor, 2001; Bohme and Cross, 2002). Blast search showed that Y71G12B.16 has homologs in sea urchins, tunicates, mollusks and vertebrates, but not in Drosophila. The vertebrate Y71G12B.16 homologs belong to the RGM (repulsive guidance molecule) family that includes RGMa, RGMb (DRAGON), RGMc (hemojuvelin) and the recently identified RGMd (Camus and Lambert, 2007; Corradini et al., 2009). Y71G12B.16 appears to be a distant member of the RGM family (Camus and Lambert, 2007), sharing 22%, 21% and 18% identity to human RGMb, RGMa and RGMc, respectively, in the region flanked by the N-terminal signal peptide and the C-terminal pro-peptide (Fig. 3B). In particular, Y71G12B.16 and human RGMb share 46% identity in the vWF type D domain and 40% identity in the hydrophobic region. Like RGMb, Y71G12B.16 lacks the RGD (Arg-Gly-Asp) motif found in RGMa and RGMc proteins (Fig. 3B). We therefore named Y71G12B.16 DRAG-1. Like C. elegans, other invertebrates including sea urchins, tunicates and mollusks also contain a single RGM-related protein in each genome (Camus and Lambert, 2007).
RGM proteins can function as BMP co-receptors (Babitt et al., 2005; Babitt et al., 2006; Samad et al., 2005). Our genetic evidence described above suggests that DRAG-1 might also function as a co-receptor in the Sma/Mab pathway in C. elegans.
drag-1 is expressed in the same cell types as the Sma/Mab pathway type I receptor sma-6
To test the hypothesis that DRAG-1 might function as a co-receptor for the Sma/Mab pathway, we first asked whether drag-1 is expressed in the same cells that express core components of the Sma/Mab pathway. A drag-1 genomic construct (pDS27), which contains the entire drag-1 genomic region including 4 kb 5′ sequences and 1 kb 3′ sequences, fully rescued the body size defects of both drag-1(jj4) and drag-1(tm3773) mutants (Fig. 3C,D). Similarly, a drag-1 cDNA construct (pCXT15; see Materials and methods), with the genomic coding region of drag-1 replaced by drag-1 cDNA and the drag-1 3′UTR replaced by unc-54 3′UTR, also rescued the body size defects of drag-1(jj4) mutants (Fig. 3C,D). These results suggest that the regulatory elements required for drag-1 function in regulating body size probably reside in the upstream sequences used in pCXT15. We then generated a transcriptional drag-1p::gfp fusion pDS15 using these upstream sequences (see Materials and methods). Transgenic lines carrying pDS15 showed GFP expression in pharyngeal, hypodermal and intestinal cells (Fig. 4A-C), the same cells that express sma-6 in hermaphrodites (Krishna et al., 1999; Yoshida et al., 2001). In fact, transgenic lines carrying both drag-1p::gfp and sma-6p::mRFP showed co-localization of the two reporters in all three tissues described above (Fig. 4A-C,E-G,I-K). Therefore, drag-1 is expressed in the same cell types as sma-6 in hermaphrodites. However, drag-1p::gfp is not expressed in the male tail cells that express sma-6p::mRFP (Fig. 4D,H,L), consistent with the lack of male tail defects in drag-1(jj4) mutant males.
The same expression pattern of drag-1 was also observed using a functional translational DRAG-1::GFP fusion (pJKL849), which has GFP inserted right before the putative cleavage site upstream of the C-terminal pro-peptide sequence (Fig. 3B). This reporter is fully functional as it rescued both the body size and the SOSMLP of jj4 mutants (Fig. 3D; Fig. 5D,O). Just like the drag-1p::gfp transcriptional fusion, DRAG-1::GFP was present in pharyngeal, hypodermal and intestinal cells (Fig. 4M-O). Robust expression in the M lineage was also detected from the 1-M to 4-M stage (Fig. 4Q-S). DRAG-1::GFP signal was reduced (40%, n=20) or undetectable (60%, n=20) at the 8-M stage (Fig. 4T), and completely undetectable beyond the 8-M stage (Fig. 4U; data not shown). The M lineage expression pattern of drag-1 is consistent with previous findings that sma-9 functions prior to the 8-M stage for proper M lineage development (Foehr et al., 2006).
drag-1 functions in the same cells as the Sma/Mab pathway receptors and SMADs to regulate body size and mesoderm patterning
The Sma/Mab pathway receptors and SMAD proteins have been shown to function in the hypodermal cells to regulate body size (Yoshida et al., 2001; Wang et al., 2002). Because drag-1 is expressed in the same tissues as sma-6 in hermaphrodites, we tested whether drag-1 also functions in hypodermal cells to regulate body size. We forced the expression of drag-1 cDNA in pharyngeal muscles [using the myo-2 promoter (Okkema et al., 1993)], intestine [using the elt-2 promoter (Fukushige et al., 1998)], and in hypodermal cells [using both the elt-3 and rol-6 promoters (Kramer and Johnson, 1993; Gilleard et al., 1999)], and tested the ability of each transgene to rescue the small body size of jj4 mutants. As a control, drag-1 cDNA under the control of its own promoter could rescue the small body size phenotype of jj4 mutants (Fig. 4P). As shown in Fig. 4P, forced expression of drag-1 in hypodermal cells rescued the small body size phenotype of jj4 mutants, whereas expression in the pharynx and intestine did not. Thus, drag-1 functions in the same cells as the Sma/Mab receptors and SMAD proteins to regulate body size.
Previous studies have shown that sma-9 and the Sma/Mab pathway function within the M lineage to regulate dorsoventral patterning of the M lineage (Foehr et al., 2006). We therefore forced the expression of drag-1 within the M lineage in drag-1(jj4); sma-9(cc604)double mutants using the hlh-8 promoter (Harfe et al., 1998). This M lineage-specific expression of drag-1 (using pCXT148) is sufficient to rescue the SOSMLP phenotype of jj4 mutants (24.5%, n=106), i.e. to reverse the M lineage phenotype of drag-1(jj4); sma-9(cc604) mutants to that of sma-9(cc604). Thus, drag-1 also functions in the same cells as sma-9 and the Sma/Mab pathway to regulate M lineage development.
DRAG-1 localizes to and functions at the cell membrane
The translational DRAG-1::GFP reporter described above allowed us to examine the sub-cellular localization pattern of DRAG-1. As shown in Fig. 4M-O and Fig. 5B-C, DRAG-1::GFP was present outside of the nucleus but localized to the cell surface as well as inside the cell. To determine if DRAG-1 protein is membrane-associated, we performed cell fractionation experiments to separate the soluble fraction and the membrane fraction using lysates from mix-staged worm populations (see Materials and methods). As controls, we used worms expressing GFP alone under the control of the hmt-1 promoter, and worms expressing the HMT-1::GFP fusion under the control of the hmt-1 promoter (Schwartz et al., 2010) (see Materials and methods). HMT-1 is a transmembrane half-molecule ATP-binding cassette transporter required for heavy metal detoxification (Vatamaniuk et al., 2005). As shown in Fig. 5N, GFP was only detected in the soluble fraction, whereas HMT-1::GFP was only detected in the membrane fraction. In the same experiment, DRAG-1::GFP was only detected in the membrane fraction. This result, in combination with results from immunostaining, demonstrates that DRAG-1 is a membrane protein, as predicted for a putative GPI-anchor protein.
To determine whether DRAG-1 functions at the cell membrane, we generated transgenic lines expressing DelC::GFP, which has the C-terminal pro-peptide deleted from DRAG-1 (Fig. 5A). DelC::GFP showed reduced signal at the cell surface (Fig. 5E,F) and became partially soluble in cell fractionation experiments (Fig. 5N). We then checked the functionality of DelC::GFP and found that it could rescue both the body size and the SOSMLP phenotypes of drag-1(jj4) mutants, but the rescuing efficiency was reduced when compared with wild-type DRAG-1::GFP (Fig. 5D,G,O). These data together suggest that the putative C-terminal pro-peptide is important, but not essential, for DRAG-1 membrane localization and function.
The membrane localization and function of DRAG-1::GFP and DelC::GFP appeared to require that the proteins enter the secretory pathway via their N-terminal signal peptide. DelN::GFP and DelNC::GFP with their N-terminal signal peptide deleted (Fig. 5A) failed to rescue the drag-1(jj4) mutant phenotypes (Fig. 5J,O) and showed a drastically reduced level of expression (Fig. 5H,I,N), possibly due to the mis-targeting and subsequent degradation of the mutant protein.
To directly test whether DRAG-1 functions at the cell membrane, we replaced the putative C-terminal pro-peptide sequence of DRAG-1 with the transmembrane domain of a well-characterized transmembrane protein LIN-12, LIN-12TM::GFP (Fig. 5A). LIN-12TM::GFP was solely detected in the membrane fraction (Fig. 5N), showed cell surface localization (Fig. 5K,L) and fully rescued the body size and SOSMLP phenotypes of drag-1(jj4) mutants (Fig. 5M,O). Thus, DRAG-1 is not only present, but also functions, at the cell membrane.
DRAG-1 positively modulates Sma/Mab signaling as indicated by a Sma/Mab-responsive reporter
The phenotypes of drag-1 mutants described above suggest that DRAG-1 positively modulates Sma/Mab signaling. To test this hypothesis, we generated a Sma/Mab-responsive reporter as there are no existing phospho-SMAD antibodies or appropriate SMAD::GFP reporters or anti-SMAD antibodies that allow us to directly monitor the output of Sma/Mab signaling.
Previous work has shown that individual SMAD complexes are capable of binding to three abutting SMAD boxes (or SMAD binding site GTCT) with high affinity (Johnson et al., 1999). Furthermore, abutting SMAD boxes organized in the RLR orientation (R, rightward; L, leftward) are bound by SMAD complexes with higher affinity than those organized in the RLL or RRR orientation (Johnson et al., 1999). We decided to place multiple copies of the SMAD boxes organized in the RLR orientation upstream of the minimal pes-10 promoter and gfp (Fig. 6A) and tested whether the expression of the gfp reporter is responsive to Sma/Mab signaling. We named the reporter as RAD-SMAD (reporter acting downstream of SMAD). The expression pattern of the gfp reporter was examined using two different integrated transgenic lines carrying RAD-SMAD (see Materials and methods), which showed similar expression patterns. We detected GFP signal from late embryogenesis until adulthood. GFP signal was detected in intestinal and hypodermal cells, including those from the P-lineages (Fig. 6B) and cells in the male tail (data not shown). The expression level of the reporter appeared rather dynamic during development, with stronger intestinal expression in embryos and L1s, but fainter in adults, and stronger hypodermal expression in L2 and L3 stages (data not shown). No gfp expression was detected in dauer animals either in the wild-type background or the daf-7(e1372) background (data not shown).
To test the specificity of the RAD-SMAD reporter, we generated two reporters carrying different mutations in the SMAD boxes, BAD-SMAD1 and BAD-SMAD2 (bad reporter of SMAD) (Fig. 6A). Transgenic worms carrying either reporter showed no gfp expression in any cells (Fig. 6C,I; data not shown). This result indicates that the SMAD boxes are directly responsible for RAD-SMAD reporter expression.
To test if the expression level or pattern of the RAD-SMAD reporter are regulated by Sma/Mab signaling, we crossed the two integrated RAD-SMAD reporters (jjIs2433 and jjIs2436) into different Sma/Mab pathway mutants and saw similar results with both reporters. As shown in Fig. 6D,E and 6G,H, RAD-SMAD expression in all cell types was downregulated in dbl-1 and sma-3 null mutants, but upregulated in lon-2 null mutants. In fact, quantification of the GFP signal intensity in various mutants correlated well with the body sizes of each mutant (Fig. 6J, compared with the body size of each mutant in Fig. 2). These results demonstrate that RAD-SMAD positively responds to Sma/Mab signaling.
As shown in Fig. 6F,J, RAD-SMAD expression was significantly downregulated in drag-1(jj4) mutants. Furthermore, the degree of reporter downregulation in jj4 mutants was smaller than that in sma-3(jj3) and dbl-1(wk70) mutants, again correlating very well with the body sizes of these mutants. These observations indicate that DRAG-1 is a positive regulator of the Sma/Mab pathway.
DRAG-1 is a cell type-specific modulator of BMP signaling in C. elegans
Previous studies have shown that RGM proteins can enhance BMP, but not TGFβ, signaling in tissue cultures by binding to selected BMP molecules as well as type I and type II BMP receptors (Babitt et al., 2005; Babitt et al., 2006; Samad et al., 2005). However, except for RGMc, there is no evidence showing that RGMa and RGMb are essential for regulating BMP signaling in vivo (Babitt et al., 2005; Babitt et al., 2006; Babitt et al., 2007; Samad et al., 2005; Xia et al., 2008; Xia et al., 2010; Andriopoulos et al., 2009). Our data demonstrate that the sole RGM homolog DRAG-1 is an integral member of the BMP-like Sma/Mab pathway in C. elegans: (1) drag-1 mutants share similar body size and SOSMLP phenotypes as mutants in all the core members of the Sma/Mab pathway; (2) DRAG-1 is a membrane-localized protein that functions at the ligand-receptor level in the Sma/Mab pathway to regulate body size; and (3) drag-1 is expressed and functions in the same cells as the receptors and SMAD proteins of the Sma/Mab pathway to regulate body size (in hypodermal cells) and mesoderm patterning (in the M lineage). Both the mutant phenotypes and the reduced level of Sma/Mab pathway reporter (RAD-SMAD) expression in drag-1 loss-of-function mutants suggest that DRAG-1 positively regulates Sma/Mab signaling in C. elegans. DRAG-1 appears to function specifically in the BMP-like Sma/Mab pathway, but not the TGFβ-like dauer pathway, as drag-1 mutants do not have any defects in dauer formation. The genetic interaction between drag-1(jj4) and certain daf-7 and daf-1 alleles (Table 1) could be owing to the fact that both the Sma/Mab pathway and the dauer pathway share the same type II receptor DAF-4 and thus exhibit low levels of crosstalk. Consistent with this notion, previous studies have also shown genetic interactions between sma-6 and daf-7 and daf-1 mutations (Krishna et al., 1999).
Interestingly, the function of DRAG-1 in modulating Sma/Mab signaling is cell-type specific. Like other core members of the Sma/Mab pathway, drag-1 functions in both body size regulation and mesoderm patterning. However, drag-1 is not expressed in the male tail and drag-1 mutants do not have any male tail patterning defects. It has been previously suggested that body size and male tail development require different Sma/Mab signaling thresholds, with the male tail requiring a lower level of signaling activity, because a hypomorphic sma-6(e1482) allele has small body size but wild-type male tail morphology (Krishna et al., 1999). Our studies on drag-1 are consistent with this hypothesis and further demonstrate that drag-1 functions to augment the level of Sma/Mab signaling activity in the hypodermal cells to ensure proper body size regulation.
RAD-SMAD, a Sma/Mab-responsive reporter in C. elegans
The activity of TGFβ signaling can be monitored by the phosphorylation or nuclear entry of SMADs, or the expression of direct TGFβ downstream genes (Schmierer and Hill, 2007; Wrighton et al., 2009). However, there are no existing phospho-SMAD antibodies, appropriate SMAD::GFP reporters or anti-SMAD antibodies, or direct Sma/Mab target genes that allow us to directly monitor the activity of Sma/Mab signaling in C. elegans. In this study, we generated a Sma/Mab signaling reporter RAD-SMAD that appears to directly reflect Sma/Mab signaling activity in C. elegans: (1) the reporter is active in cells that have been reported to be responsive to Sma/Mab signaling; (2) mutations in the SMAD binding sites abolished reporter expression; and (3) the level of RAD-SMAD reporter expression correlated with the level of Sma/Mab signaling in various Sma/Mab pathway mutants. The RAD-SMAD reporter appears to specifically reflect Sma/Mab signaling as no reporter expression was observed in dauer animals either in the wild-type background or the daf-7(e1372) background. We also noticed that the RAD-SMAD reporter is not expressed in the M lineage, where DRAG-1 and other Sma/Mab pathway components are expressed and function (data not shown). At present, we cannot rule out the possibility that the expression level of RAD-SMAD in the M lineage is too low to detect. An alternative explanation is that expression of Sma/Mab pathway targets in the M lineage requires additional transcriptional input. A likely candidate for the additional transcription factor is the SCHNURRI (SHN) protein SMA-9 (Liang et al., 2003; Foehr et al., 2006), as previous studies in Drosophila have identified composite SMAD-SHN binding sites that contain additional nucleotide sequences in between the two SMAD binding sites (Pyrowolakis et al., 2004; Gao et al., 2005). Despite this caveat, the RAD-SMAD reporter will be a useful tool for C. elegans researchers wishing to monitor the direct transcriptional output of the Sma/Mab signaling pathway.
In summary, our work demonstrates that DRAG-1 acts in a cell-type-specific manner to modulate the BMP-like signaling pathway in C. elegans, and establishes a direct link between RGMb proteins and BMP signaling in vivo. Because RGM proteins are not present in Drosophila, our work further provides a simple genetic system for mechanistic studies on RGM protein regulation of BMP pathways in vivo.
We thank the C. elegans Genetics Center, Dave Pryune, Shohei Mitani for strains and primers; Aalia AlBarwani, Rachel Fairbank and Amanda Lindy for help with the initial genetic analyses of drag-1(jj4); Ping Wang for help with statistical analysis; and Nirav Amin, Richard Padgett and Mariana Wolfner for helpful discussions and critical comments on the manuscript. This work was supported by NIH R01 GM066953 (to J.L.). Y.P. was a Howard Hughes Undergraduate Research Scholar and a recipient of the CALS Charitable Trust Undergraduate Research Grant at Cornell University. Deposited in PMC for release after 12 months.
Competing interests statement
The authors declare no competing financial interests.


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Despite all the blood and violence, none of it is your doing in this new stealth adventure – you're just the one cleaning it up.
Video game fashions come and go, but we think it's fair to say that stealth gameplay is in something of a lull at the moment. After Thief: The Dark Project and Metal Gear Solid rose to fame in the late '90s you couldn't move without a hatefully dull stealth level being shorehorned into even the most inappropriate game. That led to everyone getting heartily sick of the idea, but also highlighted that good stealth gameplay is difficult to get right – and so now something of a lost art. But thankfully it's one that Serial Cleaner still remembers.
Despite a completely different setting, Serial Cleaner has a lot in common with the early Metal Gear games. The top-down viewpoint and lack of high-tech gadgets means you're back to memorising guard routes and ducking into hiding spots when noticed. You've no camo cloaks or robot helpers to fall back on, just the knowledge that the levels are never that long and you'll instantly get the chance to try again if you're caught.
As you might gather you're not fighting cyber-terrorists or falling in love on the battlefield in this game, but instead you're cast as a 'cleaner'. Somebody that goes in after a murder and cleans up all the evidence and removes the bodies. You might have thought the cops would be a bit more careful about guarding the crime scenes once they realised you were about, but Serial Cleaner is, thankfully, far less serious about its setting than Metal Gear.
The game is set in the '70s, for seemingly no more reason than Polish developer iFun4ForAll like the aesthetic and thought it would be a good inspiration for both the funk soundtrack and the attractively angular art style. It's also an excuse not to overcomplicate your toolset, and the only thing you take with you other than your wits is a vacuum cleaner to clear up all the blood. Which we're pretty sure is not how vacuum cleaners work, unless it's supposed to be one of those heavy-duty ones you rent from Sainsbury's.
Although the details vary on each level, as a default you have to collect a number of bodies from under the noses of the law and bring them back to your station wagon parked on the edge of the map. There'll also be at least one piece of evidence you need to collect, which you end up taking home as a souvenir, and an option extra outfit or other unlockable should you want to go out of your way to pick it up.
The guards' artificial intelligence is extremely limited, as they walk along on a strict pathway and only alter their route if they notice something has gone missing. Their field of vision is clearly shown on-screen, theoretically giving you plenty of time to either run outside of its range or to hide in a nearby closet or skip. But as it turns out the police are even more stupid than they appear, and will instantly forget about you even if you jump into a hiding spot right in front of them.
As the game continues the guards become more numerous and the maps larger, forcing you to constantly improvise rather than working out a plan at the start and just trying to carry it out. We were often reminded of the now largely forgotten Commandos franchise, although this lacks that game's more complex range of abilities. It does share its difficultly though, and Serial Cleaner is certainly not a game for the impatient. Especially as the position of the bodies and evidence randomly changes every time you lose.
But if you're a fan of stealth games a certain level of difficulty is always desirable, and there's also an extra range of challenge stages where the cleaner is supposed to be drunk or you can't see the guards' vision cones. On top of that are some neat bonus levels based on classic movies ranging from Taxi Driver to Star Wars and Alien.
Much like the recent GoNNER, Serial Cleaner's biggest flaw is that it lacks that extra edge to turn it into an unequivocal recommendation. For that it need an additional gameplay mechanic or twist – some new element added to later stages that would give it extra depth and variety. Better artificial intelligence would have been enough, but as it is the game never really evolves beyond the experience presented in the first couple of hours.
That's a shame but, again like GoNNER, the low price point deflects much of the disappointment and Serial Cleaner ends up performing its role of old school stealth game with considerable aplomb. Although perhaps it's fitting that it's set in the past, because the one thing it doesn't do is move the genre forward.
Serial Cleaner
In Short: A simple, economical throwback to the golden age of stealth games, with fun presentation but a notable lack of depth and variety.
Pros: The basic mechanics work very well and the lack of complications is largely a positive. Great visual style and excellent period soundtrack. Some fun extras and bonus levels.
Cons: There's nothing about the gameplay that is very original, and the almost non-existent AI limits the experience. Later levels lack variety.
Score: 7/10
Formats: PlayStation 4 (reviewed), Xbox One, and PC
Price: £11.99
Publisher: Curve Digital Games
Developer: iFun4ForAll
Release Date: 11th June 2017
Age Rating: 16
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subset: commoncrawl


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If you've been hunting for some time, you know the significance of the right hunting boots for a successful hunt. They keep you warm in the cold, protect your feet from the rocky ground and keep your feet dry in rain. In this article, we have shortlisted the Top 7 Best Hunting Boots for ELK in order to narrow down your options.
Table of Contents
Best Hunting Boots for ELK 2021 Reviews:
Top 7 Best Hunting Boots for ELK 2021 Comparison Table:
1. Irish Setter Men's 860
Irish Setter Men's 860
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2. Kenetrek Men's Mountain
Kenetrek Men's Mountain
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3. LaCrosse Men's Alphaburly Pro
LaCrosse Men's Alphaburly Pro
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4. Muck Boot Men's Arctic Pro
Muck Boot Men's Arctic Pro
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5. Rocky Men's Bearclaw 3D
Rocky Men's Bearclaw 3D
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6. Lacrosse Men's Quick Shot 8
Lacrosse Men's Quick Shot 8
Check On Amazon | |
7. KAMIK Nationplus Boots
KAMIK Nationplus Boots
Check On Amazon |
1. Irish Men's 860 – Best Big Game Hunting Boots for Elk:
Our most favourite elk hunting boots are the Irish Setter Men's 860 Elk Big Game Hunting Boots. The Irish setter is a major brand in the world of hunting boots and these boots live up to the company's name. They are made using 100% leather which says a lot about their amazing quality and durability.
They have a synthetic sole for added protection. The outsole is built such that it keeps your feet comfortable on the rugged ground and provides good traction. The multidirectional lugs in addition to the steel shank help provide stability as well as balance while you're out hunting.
The Irish Setter Men's 860 Elk Big Game Hunting Boots also take into account foot odour which is caused due to hunting in warm weather while wearing closed boots. These boots use ScentBan technology which helps eliminate bad odour by killing the bacteria on your feet. Gore-tex lining in the boots makes them waterproof so that your feet can stay dry. The 1000g 3M Thinsulate helps provide insulation to your feet so you can stay warm and comfortable. These boots have a lace-up design which aligns them to your feet and ankles so that they stay in place all day.
One issue with these boots is that they are a little heavy which might be a little uncomfortable for beginners.
We are satisfied with the amazing quality of the Irish Setter Men's 860 Elk Big Game Hunting Boots and we would recommend them for elk hunting.
- Colour: Brown
- Dimension: 17.8 x 16.1 x 6 inches
- Material: Leather
- Breathable: Yes
- Insulated: Yes
- Waterproof: Yes
- Design: lace-up
- 100% leather
- Synthetic sole
- Lace-up design
- 1000g 3M Thinsulate insulation
- Gore-tex lining
- ScentBan technology
- Multidimensional lags
- Odour free
- Waterproof
- Insulated
- Good traction
- Heavy
2. Kenetrek Men's – Mountain Extreme Ni Hunting Boots:
The Kenetrek Men's Mountain Extreme Ni Hunting Boots are built from pure leather to make them last longer. The durable rubber sole helps provide greater traction and stability on rough terrains. The upper is built from a one-piece vamp so it does not have seams going down the tongue. This makes the boots more waterproof in addition to being resistant to abrasion. You can walk through puddles or streams and your feet will still remain dry. Resistance to abrasion makes them last longer even if you take long trips across rocky terrain.
With a Windex membrane, the Kenetrek Men's Mountain Extreme Ni Hunting Boots are extremely breathable and flexible. This also makes them more waterproof. They also feature k-talon outsoles which help in achieving a greater grip over slippery or rocky terrain. In these boots, you can climb hills and walk over streams while staying comfortable.
However, you will need to wear these shoes for a while before taking them out to hunt because they require breaking in.
All in all, the Kenetrek Men's Mountain Extreme Ni Hunting Boots provide all the right facilities required for elk hunting.
- Colour: Brown
- Weight: 3.9 lbs
- Dimension: 13.54 x 13.46 x 4.65 inches
- Material: Leather
- Breathable: Yes
- Waterproof: Yes
- Design: Lace-up
- Made from pure leather
- Rubber sole
- Upper made from one-piece vamp
- Windtex membrane
- K-talon outsoles
- Durable
- Waterproof
- Flexible
- Need breaking in
3. LaCrosse Men's Alphaburly – Best Pro Hunting Boots:
Next up, we have the LaCrosse Men's Alphaburly Pro 18″ 800G Hunting Shoes. We are a great fan of the quality of these boots and the fact that they are made pure rubber and neoprene which makes them completely waterproof. The rubber sole on these boots is very durable and delivers maximum flexibility for trekking on steep and rocky ground.
They feature a 3.5 mm neoprene core over which a layer of premium rubber has been hand-laid for maximum precision of design. This makes the boots not only waterproof and flexible but also scent-free so that your feet can remain fresh and odour free even after hunting for long hours.
The LaCrosse Men's Alphaburly Pro 18″ 800G Hunting Shoes have multiple layers of rubber around the toes which protects your toes from hitting on hard rocks and provides maximum comfort. An embossed liner on the inside of the boots helps make air circulation better to make your boots more breathable. This also wicks moisture and allows for quick drying of your boots. They have an active fit design along with an adjustable neoprene gusset which makes it easier to put your shoes on and take them off quickly.
We don't like the fact that the heels quickly lose their finish after being used for a few times.
Despite that, we are satisfied with the waterproof and flexible design of the LaCrosse Men's Alphaburly Pro 18″ 800G Hunting Shoes and we would recommend them.
- Colour: Various
- Dimension: 13 x 6 x 4 inches
- Material: Rubber, neoprene
- Breathable: Yes
- Waterproof: Yes
- Design: Active Fit
- Rubber sole
- Made of rubber, neoprene
- 3.5 mm neoprene core
- Hand-laid layer of rubber
- Multiple layers of rubber around the toes
- Embossed liner
- Adjustable neoprene gusset
- Active fit design
- Scent free
- Waterproof
- Comfortable
- Heel loses finish quickly
4. Muck Boot Men's – Arctic Best Snow Hunting Boots:
The Muck Boot Men's Arctic Pro Snow Boots are made from premium neoprene. They also have a rubber sole which makes them flexible enough for walking on rocky terrains. Shaft is around 15.25 inches from the arch and the boot opening has a circumference of 17.5 inches. 8 mm neoprene makes these boots quite comfortable by adjusting to the shape of your feet in order to prevent blisters and chaffing. This also increases their heat retention, shock absorption as well as waterproofing.
The Muck Boot Men's Arctic Pro Snow Boots are one of the warmest boots on the market owing to an additional fleece lining. There is also a 2 mm thermal foam underneath the footbed. These features make the boots comfortable and warm. These boots can keep you warm in temperatures as low as -60 degrees Fahrenheit.
Elk hunting is usually carried out in cold areas and hence these boots are perfect for this task. Stretch fit design of these boots not only keeps seals in your body heat but also helps keep debris out. In addition, they also have double reinforcement in the heels, instep and Achilles because these areas need to support the most.
One issue with these boots is that they have low breathability so in slightly warm weathers you may face sweating and bad odour issues. The Muck Boot Men's Arctic Pro Snow Boots provide warmth and comfort for hunting in cold areas and are definitely among the best hunting boots for elk hunting.
- Colour: Mossy Oak Country
- Opening: 17.5 inches
- Shaft: 15.25 inches from arch
- Material: Rubber, neoprene
- Waterproof: Yes
- Design: Stretch-fit
- Insulated: Yes
- 8mm neoprene
- Rubber sole
- Fleece lining
- 2 mm thermal foam under footbed
- Double reinforcement in the heels, instep and Achilles
- Warm
- Waterproof
- Good support
- Flexible
- Low breathability
5. Rocky Men's Bearclaw – Best 3D LTT Hunting Boots:
Rocky is a brand well known for its hunting boots. The Rocky Men's Bearclaw 3D LTT Hunting Boots have a full leather construction which makes them strong and durable. The rubber sole is sturdy and flexible and does not easily break on rough terrain. The shaft of these boots measures around mid-calf from the arch. They are one of the most highly-rated boots due to the durability they offer. The design of these boots not only provides support and protection but also better ventilation and breathability.
The Rocky Men's Bearclaw 3D LTT Hunting Boots have Thinsulate insulation along with an exclusive Rocky footbed which helps provide better air circulation within the boots. Good insulation keeps your feet warm in the cold weather and good ventilation ensures that your feet don't get sweaty and develop a bad odour. The best feature of these boots is the 3D outsole which is basically protruding lugs which help provide better traction over the slippery ground.
However, these boots are not suited for heavy-duty use so you should keep that in mind before taking them on a hunting trip.
The Rocky Men's Bearclaw 3D LTT Hunting Boots are great elk hunting boots and we recommend them.
- Colour: Black, brown
- Dimensions: 8 x 4 x 12 inches
- Material: Leather
- Waterproof: Yes
- Design: Lace-up
- Insulated: Yes
- Breathable: Yes
- Full leather construction
- Rubber sole
- Thinsulate insulation
- Exclusive Rocky footbed
- 3D outsole
- Good traction
- Insulated
- Good ventilation
- Durable
- Not for heavy duty use
6. Lacrosse Men's Quick Shot 8 – Realtree Xt:
In 6th place, we have the Lacrosse Men's Quick Shot 8 Realtree Xt Green-M which are one of the most affordable hunting boots on the market. This means that you can buy them even on a low budget. However, this does not mean that the company has compromised on the quality.
Made from 100% leather and nylon, these boots offer a reliable build and good quality. Breaking into these boots is very easy and they quickly adopt the shape of your feet. They feature 900 denier nylon which is rugged and lightweight and also a waterproof suede upper. This makes the boots lightweight and also waterproof.
Lace to toe design of the Lacrosse Men's Quick Shot 8 Realtree Xt Green-M makes them easy to wear and also cosy and comfortable. The EVA insole provides added comfort and stability. It is also removable which makes the maintenance easier. A fully waterproof dry lining keeps your feet dry and odour free by preventing moisture in your feet. They also have a rugged lug outline with a non-loading performance that helps achieve a better grip on rocky and slippery ground.
However, these boots come in slightly smaller sizes so you might have difficulty finding the right size if you have a bigger foot size. The Lacrosse Men's Quick Shot 8 Realtree Xt Green-M are decent and affordable hunting boots and you should consider investing in them.
- Colour: Brown/Green
- Dimensions: 12 x 8 x 4 inches
- Material: Leather and nylon
- Waterproof: Yes
- Design: Lace to toe
- Breathable: Yes
- 900 denier nylon
- Waterproof suede upper
- EVA insole
- Fully waterproof dry lining
- Rugged lug outline with non-loading performance
- Affordable
- Easy to break in
- Lightweight
- Waterproof
- Sizing is an issue
7. KAMIK Nationplus Boot – Best Value:
The final product of this list is the KAMIK Nationplus Boots. They are widely used by elk hunters owing to their waterproofing technology, high shaft as well as rugged design. They are built using 100% leather which makes them sturdy and durable. They provide the perfect ankle support as their high shaft is around 10 inches from their arch. They also feature a lacing system that is rustproof which keeps them new and intact for a very long time.
The KAMIK Nationplus Boots have a 0.75 inches platform. The 200B Thinsulate insulation in these boots keeps you warm even in temperatures below -40 degrees Fahrenheit. This means you can easily hunt in extremely cold areas. The rubber sole of these boots extends over the toes to provide added protection and comfort. Your feet will remain dry and odour free due to the moisture-wicking lining in these boots which prevents sweating.
One issue with these boots is that the gore-tex lining is missing due to which they are not fully waterproof. We like the KAMIK Nationplus Boots for their comfortable and stable design and we recommend them.
- Colour: Various
- Dimensions: 0.39 x 0.39 x 0.39 inches
- Material: Leather
- Design: Lace-up
- Breathable: Yes
- Insulated: Yes
- High shaft is around 10 inches from their arch
- 100% leather
- Rubber sole
- 0.75 inch platform
- Rust-proof speed lacing system
- 200B Thinsulate insulation
- Moisture-wicking lining
- Well insulated
- Good ankle support
- Durable
- Keep your feet dry
- No gore-tex lining
Final Verdict:
Good hunting boots lead to a successful hunting trip. When shopping for boots it is important to consider their features and quality. We did the research and shortlisted our favourite top 7 best-hunting boots for elk hunting. All you have to do now is take your pick keeping in mind your requirements and budget. Best of luck!
Frequently Asked Questions:
1. Are rubber boots good for hunting?
Rubber boots are good for hunting because they are waterproof and scent-free.
2. How should rubber hunting boots fit?
Rubber boots should perfectly fit your ankles and insteps but you should be able to move your toes.
3. Which is warmer neoprene or Thinsulate?
Thinsulate is warmer than neoprene.
Originally posted 2020-08-19 14:53:28.


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Your website exists to engage your audience, and if you manage your business's website, your audience is your customer. For this reason, you should always follow the question, "How often should I redesign my website?", with the question, "How satisfied am I with my website's results?"
If your website is receiving a lot of incoming traffic from Google already and you're happy with your audience engagement, then you have no need to worry about redesigning your site. However, in most cases, a redesign is necessary every two to three years. This is because improved website development helps you stay up to date with the digital revolution and can help you get the results you want. Because web design trends come and go so quickly, two or three years is all it takes for a professional, modern website to become hopelessly stale and outdated.
Here are signs your website may need some elbow grease whether or not you're currently happy with your digital marketing results:
- Your site isn't driving demand. Take a look at your website to figure out why your site isn't driving demand. In order to fix a problem, you need to find the cause. Try doing an internet marketing test called the 5-Second Test. If a new customer can understand what your business does in five seconds or less, your website is golden. If not, you may want to prioritize a redesign. Consider improving your calls to action as well so your users will know what they should do with the information you provide.
- Your web design isn't optimized for mobile users. Mobile friendliness isn't just a convenient way to improve your business' reputation among users. Up to 48% of users report assuming a business doesn't care when their website doesn't work well on mobile devices. What's more is that mobile friendliness has been an expanding feature in recent Google updates meaning a poor mobile site could be affecting your SEO.
- Your website doesn't update well. A website that routinely updates with fresh and related content is more likely to be picked up by the Google crawl bot. In terms of content updates, the answer to the question "How often should I redesign my website?" should be "frequently." If your website is difficult to update, you probably have a significant number of unrelated articles and outdated information lurking on your website.
It may be time to hire professional website design services when you find yourself asking "How often should I redesign my website?" Fortunately, MRN Designs can give you the custom web design and digital marketing services you need all in one place. Whether it's improving your Google Adwords, improving web optimization, or completely revamping your mobile responsiveness, MRN Designs can provide you with what you need. Get in touch with us today to get your digital marketing project started.


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Do not assume that every extension that sounds useful is useful. Take PDFConverterSearch4Free as an example. The name of this extension might seem promising, and the information that can be found next to its installer on the Chrome Web Store suggests that it is a beneficial tool that can optimize your web searches. Needless to say, that is usually enough for Chrome users to want to try something out. The thing is that downloading something for free is not always an inconsequential thing. Some extensions and apps can look harmless, but turn out to be malicious, and that is why you need to be cautious about what you install. Are you currently worried that you have downloaded a threat? Perhaps a malware scanner identified the seemingly harmless extension as a PUP (potentially unwanted program) and recommended removal? We do recommend deleting PDFConverterSearch4Free, and you can learn all about it in this report.
Perhaps chrome.google.com/webstore/detail/pdfconvertersearch4free/jmogbdeelhhdhbcdfblpclejnlidoikf was the webpage you downloaded PDFConverterSearch4Free from. Perhaps you used an unfamiliar installer. While it is better to use the Chrome Web Store rather than unfamiliar installers that can bundle the PUP along with malware, we do not recommend installing it in any way. Some Google Chrome users might decide to install it just because portal.pdfconvertersearch4free.com offers a free file converter, but this page is not introduced by the PUP. Instead, it is feed.pdfconvertersearch4free.com that gets introduced, and it is meant to take over the default search provider of your browser. That is exactly how extensions from the PDFConverterSearch4Free family work. We cannot say which the first extension in the family was, but some of the more recent variants include HDMusicStreamSearch, GamingSearch, and ConvertItSearch. They usually have unique installers and, obviously, names, but they always act the same once installed.
Obviously, the most noticeable change that PDFConverterSearch4Free instigates is a default search provider takeover. Unfortunately, some people might not realize this, or they might forget about it, and that is easy to do since the PUP redirects to Yahoo Search. Many Chrome users might not even link the PUP with this search engine, and that is dangerous. Why? The PUP influences the results to ensure that you are shown exactly what it wants you to see. This might include sponsored third-party content. Even if it is attractive – and it should be, given that the PUP tracks your browsing history to personalize the content – you should stay away from it because you do not know what it could hide. Perhaps the injected links could represent authentic products, services, and websites, but on the flip side, you could also be exposed to virtual scams and dangerous malware. Note that schemers know what to offer and how to display it to trick you, and so you can never let your guard down. But shouldn't Yahoo Search only show links to harmless websites? Not if a third-party search engine is involved. In this situation, we recommend staying on the err of caution.
So, how will you remove PDFConverterSearch4Free? First of all, have you scanned your system? If you think that there are no threats running around, you might think that performing a full system scan is a waste of time, but it is not. A reliable scanner only takes a few minutes to inspect your system thoroughly, and once it is done, it lists all of the threats that require removal. Ideally, PDFConverterSearch4Free is the only threat that you need to delete, but you can never be sure of that unless a full system scan is performed. If you scan your system and threats are found, we hope that you can find the guides you need on this website, but you also should think if installing an automated anti-malware tool is not the better option. This tool would instantly delete all threats, and you could also forget about protection in the future as the tool would take care of it. Have any questions? Add them to the comments below.


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Utilising the Internet of Things (IoT) in Healthcare & Medical Devices
By Mi3At Mi3 we are often asked by clients to incorporate new technologies into their designs or products. One such 'new kid on the block' technology that is starting to disrupt the healthcare sector is the Internet of Things, or IoT.
Collaborating with organisations that can develop applications in this sector strengthens our offering to our own customers. In this paper, Polymorph, custom software and application developers discuss the technology trends that are currently disrupting the healthcare sector, medical devices included.


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Tag: Dr. Kevin Huffman
Doc to providers: It's time to think big
June 22, 2010Theresa Flaherty, Managing Editor
America's obesity epidemic was front and center during healthcare reform talks. It's one of the biggest drivers of healthcare costs, but until now, doctors didn't get paid to treat it, says Dr. Kevin Huffman, a bariatric specialist who recently became a consultant to Gendron, Inc. He spoke with HME News recently about how healthcare reform-and HME providers-will help these patients.
HME News: Just how many obese Americans are there?
Dr. Kevin Huffman: Between 1980 and 2000 we saw a doubling of...
Calling Daniel Webster. Put in a good word for bariatrics.
May 21, 2010Theresa Flaherty, Managing Editor
I chatted with Dr. Kevin Huffman yesterday about how health care reform will attempt to address our obesity epidemic by making sure docs get paid for treating the disease. As that happens, the docs and the patients will have conversations about the patient's needs, including bariatric durable medical equipment (you knew I was going to say that, didn't you?).
Huffman recently became a consultant to Gendron, which offers a line of bariatric DME.
Bariatric, said Huffman, will become a more common...


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Running Late, Again
The next entry in Hooked on a Feeling is still in the works. It's going to be such a good post that I cannot finish it in one night.
Sorry to disappoint.
But worry not, for the post will land tomorrow morning. Until then, please enjoy this parody of Sherlock, produced by Studio C.
See you tomorrow!


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Say 'hello' to a deep drenching of quenching hydration. Herbal Essences Hello Hydration deep moisturising shampoo and Hair Conditioner replenishes moisture to your thirsty tresses. Soak it up and flood your senses with coconut essences. Talk about beautifully moisturised locks. Use with Herbal Essences Hello Hydration Hair Conditioner.
Key Features
- Hydrating shampoo for dry hair that helps replenish your hair's much needed moisture
- 100% recyclable bottle (our cap not yet) depending on recycling facilities
- Shampoo 0% colourants, 0% paraffin, 0% silicones
- Recognised by Peta Org for our commitment to always stand for cruelty free
How to use
Simply apply shampoo generously to wet hair, massage into a creamy lather, enjoy the scent, and rinse. For more beautiful hair, use with Herbal Essences Hello Hydration Conditioner.
Hazards and Cautions
Avoid contact with eyes. If contact occurs, rinse thoroughly with water.
Important info
Country of origin: France
Aqua, Sodium Laureth Sulfate, Sodium Lauryl Sulfate, Glycol Distearate, Sodium Citrate, Cocamide MEA, Sodium Xylenesulfonate, Cocamidopropyl Betaine, Dimethicone, Parfum, Sodium Chloride, Citric Acid, Sodium Benzoate, Guar Hydroxypropyltrimonium Chloride, Glycerin, Tetrasodium EDTA, Hexyl Cinnamal, Sodium Hydroxide, Benzyl Salicylate, Propylene Glycol, Limonene, Linalool, Magnesium Nitrate, Zea Mays Silk Extract, Orchis Mascula Flower Extract, Cocos Nucifera Fruit Extract, Methylchloroisothiazolinone, Magnesium Chloride, Methylisothiazolinone


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May is Asian and Pacific Islander Heritage Month, celebrating the history and stories of Asians and Pacific Islanders in the United States. In recognition, the National Trust for Historic Preservation recently reached out to the Greenwich Historical Society to discuss the legacy of Genjiro Yeto (1867-1924), an accomplished Japanese-born painter and illustrator who played a key role in the formation of the art colony that thrived at the Holley boarding house in Cos Cob.
Visit the National Trust for Historic Preservation's site to read our interview about how the Greenwich Historical Society interprets Yeto's legacy at the Bush-Holley House, and learn details about his close relationship with Constant Holley MacRae.
The Greenwich Historical Society is proud to be part of the Historic Artist's Homes & Studios network, a program of the National Trust for Historic Preservation.
Sharing Genjiro Yeto's story with visitors is a critical aspect of our educational and interpretive program at the Greenwich Historical Society. Born in Arita, Japan to a family with ties to the porcelain painting business, Yeto came to the United States in 1890 at the age of 23 with plans to pursue a commercial career. After achieving a measure of success in business he shifted his focus to art, enrolling in 1895 as a student at New York's Art Students League.
In New York, Yeto studied under Robert Baum, John Henry Twachtman and Kenyon Cox, finding particular resonance in Twachtman's Japanese-inspired approach to landscape and composition. The summer of 1895 Yeto joined Twachtman's summer outdoor painting class in the waterside village of Cos Cob and lived in a boarding house run by the Holley family, alongside many of his fellow students.
In Cos Cob Yeto found camaraderie and lasting friendships with several American artists, including his close friend Elmer Livingston MacRae, who married Constant Holley in 1900 and made the Holley House his permanent home. Yeto was a regular presence at the Holley House for many years, and his love for both American and Japanese culture had a lasting influence on his fellow artists.
Inspired by Yeto, Constant Holley MacRae took up ikebana flower arranging, held tea ceremonies, and practiced origami, and the artists of the Colony as a whole embraced Japanese-style dress and décor, taking part in a widespread appreciation for Japanese culture that permeated American artistic society in the late-19th century.
Through the late 1890s and early 1900s Yeto found success in the United State as a watercolor painter, chiefly specializing in elegant floral studies and genre scenes, and as an illustrator of books for children and adults. His work was exhibited widely including at the New York Watercolor Club, the National Academy of Design, and the Pennsylvania Academy of the Fine Arts. In 1900 Yeto was invited to act as a cultural consultant for the original Broadway production of Madam Butterfly.
Yeto returned to Japan in ca. 1903, where he soon was married and had his first child. In the following decade he traveled between the United States and Japan several times, exhibiting paintings in America and accepting illustration work. Eventually he settled back in Japan permanently. Research on Yeto's artistic career was limited for many years by confusion about the Romanticized spelling of his name – often recorded as "Gaingero" during his lifetime. Further, for reasons not understood by his American contemporaries, his family name of Yeto was adopted as young adult in Japan; his birth name was Genjiro Kataoka, which he reverted to using again later in life.
Readers interested in learning more about Yeto's life and work may consult Susan G. Larkin's excellent book The Cos Cob Art Colony: Impressionists on the Connecticut Shore (Yale University Press, 2000).
If you are interested in receiving a Japanese-language site guide to the Bush-Holley House, prepared by Noboru Uezumi please send an email to email@example.com.
Maggie Dimock is Curator of Exhibitions and Collections at the Greenwich Historical Society.


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Member Highlight: April Odom's journey from medical assistant to nurse practitioner and business owner
Posted over 3 years ago
April Odom participated in a medical assistant program in high school, and her plan was always to go into nursing.
But then, as it often does, life happened.
Odom had a baby and her career took a wandering road. For six years, she worked as a pharmacy technician. Then she went back to school. Odom was one of 40 students accepted to Malcolm X's ADN program. She earned her CNA while in nursing school and worked in an in-patient rehabilitation unit. She stayed with the unit when she graduated from Malcolm X in 2008 and worked as a registered nurse.
Odom describes that unit as a shining example of what diversity in the nursing workforce should look like. "Because we were a rehabilitation unit, we collaborated with professionals from a variety of other professions. Beyond that, the unit itself was small but so diverse. We had people from different specialties as well as different backgrounds, nationalities and interests. We were able to learn from one another, collaborate, and take advantage of one another's different backgrounds and perspectives. Together, that allowed us to take care of the whole patient."
Mentorship helped her reach higher
Odom had a mentor on the rehab unit, and that mentor encouraged her to continue her education. She went on to earn her bachelor's degree in nursing online from Jacksonville University, and then went straight on to earn her Master's in Nursing at Lewis University.
Odom graduated as a nurse practitioner in 2016. After a year spent working in family practice, she opened a business: Physicals Plus, Health and Wellness.
"In the beginning, I continued working full-time and operated the business part-time. Now, it's more like full-time, full-time," Odom laughs. "We do health promotion, vitamin therapy, physicals, disease prevention, lifestyle-based weight management and provide house calls to the senior population. Eventually, the goal is to have a full-service family practice clinic with collaboration with multiple specialties and providers."
Odom credits ISAPN as part of what inspired her to start her own business. "I joined while in NP school after my professor showed a video about being a member. At the time, all I knew was my own tight-knit unit, and I wanted to get out of my comfort zone."
The opportunities are unlimited
Now, Odom serves as ISAPN's Vice President.
To others considering becoming an APRN or an ISAPN member, Odom says, "Don't even hesitate. The opportunities you will have, the things you can do, the places you can go, are unlimited. ISAPN brings you to the forefront of the profession, and APRNs are filling the gaps in access to care. We need you.""ISAPN has helped me grow and has taught me that together, we are a force. ISAPN fought for and achieved full practice authority for APRNs in Illinois amongst other legislative accomplishments. Being a member has shown me how when we come together, we can make change. Beyond that, being a member has connected me to a network of like-minded professionals from so many different backgrounds. That diversity of ideas and professional support empowered me to reach higher. I started as a medical assistant student and climbed every rung of this ladder one step at a time. Now, I own my own business."


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Beavertown Brewery - Gamma Ray American Pale Ale
Beavertown Brewery are based in Tottenham in London and are one of the most recognisable and well-respected craft beer brewery in the UK. They are known for their distinctive artwork and their creative beer ideas - they have even hired an in-house designer!
Gamma Ray is an APA that is tropical, juicy and with a punch-y aroma. A very good session beer.


text: Could the President's Twitter fingers pose a legal challenge? On Friday, President Donald Trump's Twitter account posted one tweet, deleted it and issued another tweet with similar language. For posterity's sake, the first tweet read: "The FAKE NEWS media (failing @nytimes, @CNN, @NBCNews and many more) is not my enemy, it is the enemy of the American people. SICK!" It was quickly replaced with: "The FAKE NEWS media (failing @nytimes, @NBCNews, @ABC, @CBS, @CNN) is not my enemy, it is the enemy of the American People!" Deleting a tweet is commonplace – Twitter does not allow users to edit their messages after they post. The question is whether Trump is violating the Presidential Records Act of 1978, which requires all the president's records be preserved for eventual release to the public on a delayed basis long after the commander in chief leaves office. The National Archives and Records Administration is tasked with managing the records. Reached by CNN on Friday, NARA spokesman John Valceanu said the PRA should be followed but directed all questions about the current president to the White House. Reached for comment, Kelly Love, a White House spokeswoman, said Saturday, "We have systems in place to capture all tweets and preserve them as presidential records; even if they have been deleted." She did not immediately provide further details, including whether those "systems" would apply to Trump's personal account as stringently as they do the @POTUS account. The PRA specifically does not distinguish between personal and work efforts, and the last provision of the law prohibits personal communications unless there is government redundancy. Guidance from NARA on the act said presidential records "can be in any media, including textual, audiovisual and electronic." This would almost certainly include tweets from the President's personal account as it does to the official @potus account handed down to Trump from former President Barack Obama. Trump often deletes tweets from his personal account over spelling mistakes and then quickly tweets corrected versions. There are caveats in the law for personal records, like a private diary, but the act "requires the president and his staff to take all practical steps to file personal records separately from presidential records." The responsibility for complying with the law lies with the President. Whether the President has violated the spirit of the law is a trickier question. "The PRA gives ownership of all presidential records to the United States (we the people), and all of those records must be transferred to the Archivist of the United States after the president leaves office," Shontavia Johnson, a law professor at Drake University Law School, wrote in an email to CNN. "The only presidential records that can be disposed of are those that 'no longer have administrative, historical, informational, or evidentiary value.' To completely dispose of presidential records, including tweets, the president has to go through certain complex steps with the Archivist, who has years of training maintaining such information." Kathleen Clark, an ethics expert and professor at Washington University Law, echoed some of Johnson's thoughts but added, "On the other hand, a government official – even the President – has the right to speak in his personal capacity about his work."


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05/11/2020 Equality and Non-Discrimination
PACE's Parliamentary Network Women Free from Violence and Committee on Equality and Non-Discrimination are organising a Facebook Live webinar entitled "Preventing and combating violence against women in sport", on 10 November at 10 am.
Speakers include former athletes and specialists in equality issues in sport, working on these issues from the Czech Republic, Ireland, France and Belgium. Moderation will be provided by Killion Munyama (Poland, EPP/CD), who is preparing a report on " The fight for a level-playing field – ending discrimination against women in the world of sport". Introductory remarks will be made by the General Rapporteur on violence against women, Zita Gurmai (Hungary, SOC).
The panelists will examine the extent of the phenomenon of violence against women in sport, the underlying causes that allow such violence to occur and the "code of silence" around its denunciation. Recommendations and good practices in force in different European countries will also be put forward.
"Gender inequality and gender-based stereotyping in sport are a mirror of the gender inequalities and discrimination present in society as a whole. The world of sport is not immune to violence against women and girls," said Mr Munyama. "It is of crucial importance that sports federations respond to claims of abuse and provide support to victims. This webinar will provide an opportunity to discuss good practices in preventing and combating violence against women in sport."
The webinar will be broadcast live on the Facebook page of the Parliamentary Network Women Free from Violence: Parliamentary Network Women Free from Violence


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Comprehensive Care Management
This idea drives our approach to care management—an approach that combines patient-centered clinical solutions, continuous improvement and
patient engagement, innovative IT applications, data analytics, and customized offerings to those responsible for the quality and cost of care.
When it comes to making a move to value, organizations face numerous challenges. ICS deploys a unique set of solutions to meet these challenges head on with an approach that matches the specific needs and opportunities of our customers and the patients and markets they serve.
Learn MoreMedicare & Dual-
Eligible Patients
Accountable Care
Integrated Delivery
Problems We Solve
Controlling patient outcomes and costs is difficult. Post-acute healthcare represents one of the fastest-growing areas of the healthcare system with the highest variation in patient outcomes and costs. ICS' risk-sharing partnership approach helps providers to better manage the following:
- Prevention of costly ED visits and hospitalizations
- Coordination with primary care physician services
- Level of care determinations
- SNF length of stay
- Factors that reduce readmissions
- Developing partnerships with high-quality skilled nursing and home health providers
- Transitioning and maintaining patients in communities where they reside and call home
- Addressing social determinants and patient support
"When you think about the health system and ICS working together to develop an end-to-end care experience for our patients, that is what value-based care is all about."
– Dr. Nihar R. Desai, Medical Director for Value Innovation, Yale New Haven Health
Our patient-centric solutions prioritize the patient experience while also maximizing efficiencies and generating total cost-of-care savings for the hospital systems and customers we serve. By combining innovative technology and robust data with compassionate and knowledgeable care, we ensure the best outcomes for patients and customers.
Learn MoreInsight
& Analytics
Care Management
& Coordination
Customized Advisory
Intuitive Technology
Improvement &
ICS' proprietary high-tech, high-touch clinical model helps organizations safely treat patients where they are most comfortable—in their home. Not only does this keep patients safe and satisfied, but it offers tangible improvements in outcomes for both patients and the providers charged with their care.
To coordinate care, and track and analyze outcome, ICS developed our own proprietary software. Care+ provides our Clinical Integration Nurses (CINs) with the tools and automation needed to deliver excellent care and provide patients with the best possible experience. This powerful software also performs the analytics, data, and reporting needed to drive measurable outcomes and control costs for our provider partners.
Driving Our Overall Method and Approach
We ensure the following:
- Patients avoid hospitalizations and ED visits by actively managing chronic conditions and comorbidities
- Coordination with patients' primary care and specialist physicians to optimally manage clinical and social needs
- Patients are treated in the most appropriate post-acute setting upon hospital discharge
- Patients are referred to the highest quality providers to meet their needs
- Patients stay for the right amount of time, receiving the right amount of services
- Everyone involved in the patient's care has the right information and incentives to achieve optimal outcomes
- Patients remain safely at home as much as possible
COVID-19 Information
Discharging patients amid the COVID-19 pandemic has many providers facing unique challenges. With a focus on quality, our patient care coordination team leverage our network of post-acute and home health solution providers to deliver excellent, CDC-compliant care and improve patient outcomes.
We are closely monitoring the Coronavirus to provide safety for the patients we serve.


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Have No Pity. Use Your Empathy And Show Compassion Instead
Little John is just learning to walk. He's making his first attempts to do this simple and so complicated action called walking. See him smiling and giggling and so faithfully making attempt after attempt to walk. He takes a few … Read the rest


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14-year-old Hayley Okines has co-written a book about her experience of living with Progeria. We talk to her mother about their story so far…
- Blog
- Published on Monday, 05 November 2012 10:00
- Last Updated on 09 November 2012
- Jasmine Joynson
- 3 Comments
Kerry Okines' daughter, Hayley, suffers from Hutchinson Gilford Progeria Syndrome. This condition means that she ages about eight times faster than the average person. I asked Kerry about the work both she and Hayley do to raise awareness of the condition and the progress that they have already made. Despite being told she would probably not make it past her thirteenth year, Hayley is fast approaching her fifteenth birthday. As well as taking part in drug trials, being the subject of articles and documentaries and getting on with life as a 14-year-old, Hayley has co-written a book with her mother.
Can you talk us through how Hayley was first diagnosed and what you were told about the condition then?
Hayley was undergoing tests for about a year. From the age of one to just before her second birthday they weren't sure what was wrong with her. They did blood tests, chromosome tests, they tested for cystic fibrosis. Because it's so rare the doctors hadn't come across it before. She was diagnosed by taking a skin sample and when they grew those cells the tests showed that the skin had very little elasticity which is something you find in older people. So then we were told it was Progeria.
You have chosen to very publicly address Hayley's condition to raise awareness. Was this a difficult decision to make?
No, at the time it felt like the right thing to do. When we were first told about the condition it knocked the whole family for six. We got a lot of positive responses from doing the media work because the condition is so rare. We had people who fund-raised to send her to Disneyland. It really just spiralled very quickly. We were in one newspaper and then invited for a magazine shoot. People were interested in making a documentary.
What would you like to see happen as a result of your work?
We've had some good results already. Hayely's been taking place in clinical trials in America which started just over four years ago. When Hayley was diagnosed there was nothing like that happening – people didn't even know which gene was linked to progeria. Now they've identified the gene and this is the forth clinical trial Hayley's been a part of. We've also helped three other families get diagnoses through doing the media. It's hard work but it's definitely been worthwhile.
Whose idea was it to write Old Before My Time?
It was a joint decision between me and Hayley. She'd done the documentary and magazine stories. Katie Price was doing documentaries and writing books so we joked that it would be funny if Hayley got her own book on the shelves. The whole thing just started from there. We've had really, really positive feedback. People are so impressed by how brave Hayley is.
You've recently had results from a study Hayley took part in. Can you outline what these drugs do and how they could be useful to people who suffer with Progeria?
These drugs are not only useful for Progeria children they will also help people that suffer with heart disease. They found some very positive results with the flexibility of the arteries in the heart. In older people, and people with Progeria, the arteries in the heart become stiffer which then causes blood flow problems which can lead to heart attacks and strokes. They've found that this new drug increases that flexibility and so lessens the risk of these. It's taken a long time but it's been worth waiting for.
Have you encountered any problems getting support for caring for Hayley? Do you think that there's anything that could be done to improve the lives of children and their families that live with serious health problems?
It was quite difficult in the beginning. We had bereavement counselling and counselling as a family. We've just had the house adapted. A lift's been put in because Hayley is in a wheelchair full-time now as her hips are permanently dislocated. Getting the funding and financial support for that was not too difficult. I think that it's really the emotional support that is sometimes harder to some by. Hayley has a visit about once a month from her friend from Demelza (which is a charity that works with children who have life-shortening illnesses). I think it's really important that she has this time so she can talk about anything she's worried about. They go out and get a coffee or a pizza and have a chat. We had a lot of support as a family but I think that it would have been nice for Hayley to have more emotional support from earlier on.
When Hayley's not travelling back and forth to the US or writing what does she enjoy doing?
She's always on Twitter. She's a little bookworm – at the moment she's reading The Hunger Games. She loves The Wanted, she's met them a couple of times and she collects all the posters. Hayley also enjoys hydrotherapy once a week. She loves sleepovers, make-overs, the kind of thing any 14-year-old likes.
If you are interested in finding out more here is a link to Amazon's page about Old Before My Time:
Image credit: photographer Chelsey Browne
Editorial Assistant at London Mums. She loves travelling, food, literature and cinema.


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Chattycast 95: Don't Worry, Loves! The Chattycast's Here!
The Chattycast gives in to one of its host's demands and devotes an entire show to Blizzard's latest.
Hey everyone. It's me, Bryan, AKA Doctor Games. I'm writing the Big Topic this week. Where's Steve, you ask? Don't worry about it. Also don't check the broom closet, no matter how much muffled yelling and clattering you hear. It's just ghosts. Ghosts who won't listen to insightful suggestions.
Anyway, since this week is the launch of Overwatch, a game I have turned a critical eye toward and found to be the single most important video game story of the year, I have declared editorial fiat in making this week's Chattycast all about Blizzard's first fully new IP in years. But rather than just spend the entire show talking about how great the game is – and I could certainly do that – I figured it'd be interesting to cast Overwatch in the larger context of the genre.
Could this be the hallmark of truly cooperative shooters? What does the popularity of a game that does not track the traditional K/D stats mean for improving the often toxic nature of online gaming communication? Are hero-based shooters in which you learn classes and characters like fighting game combatants the next big thing?
Be sure to visit AudibleTrial.com/Chattycast for your 30-day trial with a free audiobook download of your choice.
Bryan Carr posted a new article, Watch Chattycast go overboard on Overwatch at 1 PM ET / 10 AM PT


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1. Terms and Conditions of Sale:
A. CBT Supply, Inc. offers standard Terms and Conditions of Sale to approved accounts.
B. To obtain approval, commercial, business and institutional entities must provide us with a signed Business Credit Application
faxed to 800-770-7042. Contact sales or accounting to received the Credit Application by email.
C. CBT Supply, Inc. accepts all major credit cards. For security purposes, all credit card information should not be sent by email but must be sent via
fax to 410-308-4986 .
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A. We do not conduct transactions online. The information provided by web site users is for pricing purposes only. Filling out and submitting an iQuote does not obligate the person or entity to purchase anything. All information is held in strictest confidence and is not shared, rented or sold to e-mail marketers–or anyone–for any purpose.
B. Swatches shown are Web reproductions and differ from the actual product colors because every computer monitor is set differently and cannot reproduce every color exactly. CBT Supply, Inc. will not be held liable for color decisions approved solely on the basis of web color representations, and such contracts are binding based on laminate manufacturer's colors specified. For critical color decisions, please request and approve actual finish or fabric samples to complete your order. Click Here to request samples.
3. GSA Contracts:
CBT Supply, Inc. provides goods and services through the United States General Services Administration (GSA) under the following contract numbers: GS-27F-0004P (Classroom Furnishings. Details may be viewed on our GSA page.
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CBT Supply, Inc. warrants that our products are covered for defects in materials and workmanship under a Limited Warranty to the original owner of the product under normal use and conditions. Note that certain items, accessories and components by our suppliers are covered under separate warranties with varying terms and conditions. Please contact us for additional information.
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Nevertheless, CBT Supply is pleased to provide free space-planning and ergonomics information and offer it, without warranty or liability, for non-commercial use. Please inform us of your intent prior to publication.
6. MAS Green Certified Products
Materials Analytical Services, LLC has Certified SMARTdesks® for their Classroom Products, Classroom Systems, Conference Tables, Podiums, Teleconferencing, and Monitor SupportFor Product Designation as Low VOC Emitting Furniture in Accordance with the Following Standards:
• LEED for Schools
• ANSI/BIFMA e3-2012 Section 7.6.1, 7.6.2, and 7.6.3
• Collaborative for High Performance Schools (CHPS)
• California Dept. of Public Health (CDPH) Standard Method V1.1
• Criteria for the MAS Certified Green® Program
Certificate Number: MAS-1400714
Martin R. Bennett, Senior Consultant
Materials Analytical Services, LLC Certificate Valid: June 2014 to June 2015
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The SMARTdesks® organization is focused on principles of sustainability. We strive
continually to use and manage natural resources responsibly, conserve energy in our daily
operations and minimize material waste in the production of our products. Our policy is to
divert waste from the manufacturing process such that waste is either eliminated or
recycled or disposed of safely and responsibly within the scope of existing local and state
requirements and recommendations. We are dedicated to assisting our customers in
meeting the goals of sustainable-design projects and those adhering to LEED certification
guidelines. Download the complete LEED Statement pdf for your review:


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Congratulations, Susan W.!
Lincoln City Antique Week 2016 was a huge success! Not to mention a blast. We had a great time meeting folks from all over the country.
The winner of the gift basket drawing from Nelscott House Antiques and Collectibles was Susan W. We hope you enjoy your goodies.
And we're continuing to celebrate Antique Week all month long with our 30% off sale. Come on down to see what kinds of fun memories we can dig up from the past. Including this portable phonograph we recently had available, circa 1910.