text:
Published in Volume
107, Issue 5
(March 1, 2001)J Clin Invest.
Copyright © 2001, American Society for Clinical Investigation
Hepatic cholesterol metabolism and resistance to dietary cholesterol in LXRβ-deficient mice
1Department of Biosciences at Novum, Karolinska Institutet, Huddinge, Sweden
2Metabolism Unit, Center for Metabolism and Endocrinology, Department of Medicine, and Molecular Nutrition Unit, Center for Nutrition and Toxicology, Novum, and
3Division of Clinical Chemistry, Karolinska Institutet at Huddinge University Hospital, Huddinge, Sweden
4Center for Genomics Research, Karolinska Institutet, Stockholm, Sweden
Address correspondence to: Gertrud Schuster, Karolinska Institutet, Center for Biotechnology at Novum, Hälsovägen 7, S-141 57 Huddinge, Sweden. Phone: 46-8-608-9147; Fax: 46-8-774-5538; E-mail: email@example.com.
Published March 1, 2001
Submitted: March 6,
2000; Accepted: January 23,
The nuclear oxysterol-receptor paralogues LXRα and LXRβ share a high degree of amino acid identity and bind endogenous oxysterol ligands with similar affinities. While LXRα has been established as an important regulator of cholesterol catabolism in cholesterol-fed mice, little is known about the function of LXRβ in vivo. We have generated mouse lines with targeted disruptions of each of these LXR receptors and have compared their responses to dietary cholesterol. Serum and hepatic cholesterol levels and lipoprotein profiles of cholesterol-fed animals revealed no significant differences between LXRβ–/– and wild-type mice. Steady-state mRNA levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase, farnesyl diphosphate synthase, and squalene synthase were increased in LXRβ–/– mice compared with LXRβ+/+ mice, when fed standard chow. The mRNA levels for cholesterol 7α-hydroxylase, oxysterol 7α-hydroxylase, sterol 12α-hydroxylase, and sterol 27-hydroxylase, respectively, were comparable in these strains, both on standard and 2% cholesterol chow. Our results indicate that LXRβ–/– mice — in contrast to LXRα–/– mice — maintain their resistance to dietary cholesterol, despite subtle effects on the expression of genes coding for enzymes involved in lipid metabolism. Thus, our data indicate that LXRβ has no complete overlapping function compared with LXRα in the liver.
Cholesterol is a basic component of all cellular membranes and precursor of several hormones, vitamins, and bile acids, and therefore is an essential molecule for life. However, it is well established that elevated levels of cholesterol in plasma represent a risk factor for the development of atherosclerosis (1, 2).
The biosynthesis and catabolism of cholesterol are regulated by a complex network of enzymes, which are tightly controlled by several hormones (3), by cholesterol itself (4), and possibly also by oxidized derivatives of cholesterol — the oxysterols (5). Recent studies have identified several oxysterols, in particular 24(S), 25-epoxycholesterol, and 24(S)-hydroxycholesterol (6–8), as natural ligands for the nuclear liver X-receptor α (LXRα; NR1H3 from the Nuclear Receptors Nomenclature Committee, 1999) (9), suggesting that LXRα is important in cholesterol homeostasis. Mice lacking LXRα indeed lose the natural resistance to a diet rich in cholesterol, being unable to adequately regulate the conversion of cholesterol into bile acids (10).
In parallel to LXRα, another orphan receptor, LXRβ (NR1H2; from Nuclear Receptors Nomenclature Committee, 1999) (9) was cloned and characterized (11–14). While the expression of LXRα is limited to organs such as liver, intestine, kidney, adipose tissue, and adrenals (15, 16), LXRβ is expressed in all tissues thus far analyzed (11–14). LXRα and LXRβ share a high degree of amino acid similarity (78%) (17) and have thus been proposed to be paralogues. Recently, a similar function for both LXRs has been shown in the regulation of ABCA1 and ABCG1, two transmembrane proteins mediating efflux of phospholipids and cholesterol from macrophages and enterocytes (18–22). Due to the similar ligand-binding ability as well as their specificity for endogenous oxysterols, a function of both LXRs in the regulation of hepatic cholesterol and bile acid metabolism has been suggested (6, 7).
With the aim to determine if LXRβ has a regulatory function in the response to cholesterol challenge, as described for LXRα (10), mouse lines deficient for the LXRβ or LXRα genes were generated and challenged with dietary cholesterol. Our results show that, in contrast to LXRα-mutant mice, LXRβ-deficient mice maintained their natural resistance to dietary cholesterol. This indicates that LXRβ does not have a role comparable to that of LXRα in hepatic cholesterol metabolism.
Production of LXR-deficient mice. For the LXRβ-targeting vector a 6.1-kb EcoRI-ApaI genomic DNA fragment (European Molecular Biology Laboratory [EMBL] accession number AJ132602) (17), cloned into pBluescript, was used (Figure 1a). The DNA fragment between the BglII and NcoI sites, located in intron 4, was replaced by a 1.1-kb loxP cassette isolated from pGEM-30 (23). The AvrII site in intron 6 was used for insertion of a 1.4-kb SalI-XbaI loxP-flanked neo cassette isolated from pL2neo (24). The unique ApaI site at the 3′ end of the genomic insert was used for linearization of the vector before electroporation.
Strategy for the creation of LXRβ- and LXRα-deficient mice using a Cre/loxP system. (a and b) The genomic organization of LXRβ and LXRα, the corresponding targeting constructs containing the loxP sites, the predicted homologous recombinant alleles, and the LXR locus after Cre-mediated deletion are shown. Filled boxes indicate exons, arrows loxP sites, open boxes neomycin resistance and thymidine kinase genes. Filled bars indicate probes used for Southern blot analyses. H, HindIII; B, BglII; DBD, DNA-binding domain; ATG, translational start codon; TAG/TGA, stop codons. (c) Southern blot analysis. DNA was prepared from mouse tails and digested with BglII or HindIII as described in Methods. (d) Northern blot analysis. Poly A(+)-enriched RNA was extracted from a pool of livers (five females) from each respective group. Hybridization was performed by using full-length LXRα and LXRβ cDNAs as probes.
For the LXRα-targeting vector a 6.9-kb NcoI-NdeI genomic DNA fragment (EMBL accession number AJ132599) (17) was cloned into pGEM-5 (Figure 1b). The AflII site in intron 3 was used to insert a 153-bp cassette containing a single loxP site obtained from pGEM-30. A recreated HindIII site at the 5′ junction served as a restriction site for diagnostic Southern blot analysis (Figure 1, b and c). The SmaI site in intron 5 was used to insert a blunt-ended 1.3-kb XhoI fragment containing the neo cassette and a loxP site. For negative selection, a herpes simplex virus thymidine kinase cassette (HSV-tk) derived from pIC19R/MC1-TK (25) was cloned downstream of the short arm of the targeting vector. A unique BamHI site at the 3′ end of the thymidine kinase cassette was used to linearize the targeting vector.
R1 embryonic stem (ES) cells established from inbred 129/Sv embryos (26) were electroporated with 20 μg of the respective linearized targeting vector. After 48 hours, positive clones for either LXRβ or LXRα were selected with G418 or with G418 plus ganciclovir, respectively. Resistant ES cell clones were screened for hypomorphic (hyp) alleles using PCR. Homologous recombination was confirmed by Southern blot analysis (not shown). Recombinant ES-cell clones were injected into C57BL/6 blastocysts. Mating of chimeric males with C57BL/6 females led to the integration of the hypomorphic LXR alleles into the germ line. Disruption of the LXR alleles was performed by Cre-assisted deletion in vivo by mating LXRhyp/hyp and LXR+/hyp mice with cre-transgenic deleter mice (with hybrid C57BL/6-129/Sv genetic background) as described previously (27). Subsequent matings of offspring served to segregate the desired LXR+/– mutations from the cre transgene. Starting from this population of LXRα+/– and LXRβ+/– founder animals, we first crossed heterozygous LXR+/– animals and later also homozygous LXR–/– mice, as well as their respective wild-type littermates. All animals used for experiments were maximally third-generation descendants of the original heterozygous (+/–) founder animals. All mice used had a mixed genetic background based on C57BL/6 and 129/Sv strains. During establishment of all LXR-deficient mouse lines, genotypes were determined by Southern blot analysis of tail DNA (Figure 1c) and digested with HindIII (LXRα) or BglII (LXRβ). After colonies had been established, a PCR approach was used for genotyping.
Experimental procedure. Mice from the two different knockout lines and corresponding control lines (designated LXRα+/+ and LXRβ+/+), derived by intercrossing wild-type littermates for each knockout, were studied in two separate experiments performed at different times. Mice were maintained on a 12-hour light/12-hour dark cycle and fed a low-fat rodent diet (diet R36; Lactamin AB, Vodstena, Sweden) ad libitum. Twelve- to 16-week old mice were challenged with R36 chow enriched with 2% cholesterol (AnalyCen Nordic AB, Lidköping, Sweden) for 28 days. In a separate experiment, serum cholesterol and triglyceride levels from the distinct LXRα+/+ and LXRβ+/+ control mice fed standard chow for 18 months were compared. No differences were observed (data not shown). Experiments were approved by the Karolinska Institutional Animal Care and Use Committee.
Chemical analysis of serum and tissue. After 5-hour fasting, blood was drawn from mice by cardiac puncture under light methoxyflurane anesthesia. Animals were sacrificed by cervical dislocation, and tissues were immediately collected for further analyses. Blood was transferred into tubes, and serum was collected by centrifugation. Total cholesterol in serum was determined individually using a Monarch automated analyzer (ILS Laboratories Scandinavia AB, Sollentuna, Sweden) on fresh serum. Size fractionation of lipoproteins was performed on 10 μl of pooled sera from each group using a fast-performance liquid chromatography (FPLC) column (30 × 0.32 cm Superose 6B; Amersham Pharmacia Biotech, Uppsala, Sweden), and cholesterol content in lipoproteins were determined online using a commercially available reagent (Boehringer Mannheim Biochemicals Inc., Mannheim, Germany). Absorbency was monitored at 500 nM and collected using EZ Chrom software package (Scientific Software Inc., Pleasanton, California, USA). Hepatic lipids were extracted according to the methods of Folch et al. (28) and analyzed for total cholesterol using commercially available kits (Boehringer Mannheim Biochemicals Inc.). Oxysterols (24- and 27-hydroxycholesterol) in pooled serum samples were determined by isotope-dilution mass spectrometry using deuterium-labeled internal standards as described previously (29). Alanine and aspartate aminotransferase activities were measured in pooled sera according to the International Federation of Clinical Chemistry and Laboratory Medicine (30) using reagents from Roche Diagnostics (Basel, Switzerland) and a Hitachi 917 automatic analyzer (Hitachi Ltd., Naka, Japan).
Northern analysis. Northern analysis was performed on 2–10 μg poly A+ mRNA, extracted from pooled livers of five female mice from each group. The cDNA probes were generated by RT-PCR with murine RNA from liver as previously described (10, 31) or using the following primers: cholesterol 7α-hydroxylase (Cyp7a), 5′-AGATTCTCCCCTTGGGACGTTTTCCTG-3′ and 5′-AACGCTCAGCAGTCGTTACATCATCCAGT-3′; oxysterol 7α-hydroxylase (Cyp7b), 5′-GCTCCCTACAGAGCCGCCAGCTCGTC-3′ and 5′-ACGGAGCTCAGCACATGCAGCCTTACTCTG-3′; sterol 27-hydroxylase (Cyp27), 5′-GGACAGCAGTGGTACCATCTGC-3′ and 5′-CCAGTTATTCATGTATCGCTTCC-3′; farnesyl diphosphate (FPP) synthase, 5′-GGTCCTGGAGTACAACACTGTAGGAGGCAA-3′ and 5′-TGGCAGCCGCGATAGGCAGGTAGAAAGAGT-3′.
The RT-PCR products were cloned into T-tailed pBluescript plasmid (Stratagene, La Jolla, California, USA) modified as described previously (32). The sequences of the different clones were confirmed using the BigDye Terminator Cycle Sequencing Ready Reaction Kit (Perkin Elmer Applied Biosystems, Warrington, United Kingdom). Plasmids containing probes for stearoyl CoA-desaturase-1 (SCD-1), fatty acid synthase (FAS,) and sterol 12α-hydroxylase (Cyp8b1) were kindly provided by Stefan Alexson (Karolinska Institutet, Huddinge, Sweden), the probe for SREBP-2 by Christine Sadek (Karolinska Institutet, Huddinge, Sweden). Probes were labeled with [α-32P] dCTP using random prime-labeling system (Amersham Pharmacia Biotech, Uppsala, Sweden). After hybridization, signals on Northern blot filters were quantified by an image analyzer (Fuji Photo Film Co., Stockholm, Sweden) and standardized according to GAPDH mRNA expression. The data shown resulted from two or more independent determinations.
Statistics. Data are presented as mean plus SEM. The significance of differences between groups was tested by two-way ANOVA followed, when appropriate, by post hoc comparisons of group means according to honestly significant difference methods (Statistica software; Stat Soft Inc., Tulsa, Oklahoma, USA) (33). The authenticity of the absence of significant differences was determined by post hoc power calculation, when suitable.
LXR-β and LXRα-deficient mice were created according to the strategy shown in Figure 1. We first generated mutant mice by flanking two exons encoding the DNA-binding domain and the hinge region of the respective LXR genes by loxP sites (Figure 1, a and c). Mice homozygous for these hypomorphic LXRβ or LXRα alleles (hyp/hyp) were viable, and no apparent phenotype was observed. Both LXR genes were successively disrupted by in vivo Cre-mediated deletion of the loxP-flanked gene segments. Southern blot analyses proved the proper homologous recombination (Figure 1c). Northern blot hybridizations of hepatic mRNA was performed to control for the presence of the right-truncated mRNA transcripts (Figure 1d). Unexpectedly, no LXRβ transcripts were detected in LXRβ–/– mice. However, in LXRα–/– mice the expected truncated LXRα transcript lacking the 500 nucleotides, corresponding to exons 4 and 5, was formed (Figure 1d).
LXRβ–/– mice were fertile and could produce offspring. No macroscopic phenotype was observed under standard conditions.
Since LXRβ and LXRα bind the same oxysterols with similar affinity (6, 7) and since Peet et al. (10) have shown that LXRα-deficient mice lose their natural resistance to dietary cholesterol, we addressed the question of whether LXRβ-deficient mice respond to cholesterol feeding in a similar way. Male and female LXRβ–/– mice and their corresponding wild-types were challenged with 2% cholesterol-enriched diet for 4 weeks. In a separate experiment LXRα–/– and their wild-types were treated similarly.
On a standard diet, no differences in the macroscopic appearance of the livers of LXRβ–/– and wild-type controls were seen. Surprisingly, livers from LXRβ–/– mice on 2% cholesterol were also similar to those from the corresponding wild-type controls (not shown). As expected, the livers of LXRα–/– mice on 2% cholesterol showed a dramatic change in color and an increase in size (Figure 2). Chemical analysis of hepatic extracts confirmed that the physiological resistance to dietary cholesterol was preserved in LXRβ–/– mice. An increase in hepatic total cholesterol of a magnitude similar to that of wild-type controls (less than twofold) was observed in LXRβ–/– mice after cholesterol feeding (LXRβ–/– vs. wild-type, not significant; Power >95%; Figure 3a), whereas a fivefold increase was found in LXRα–/– mice (Figure 3b).
Macroscopic appearance of livers from LXRα+/+, LXRα–/–, and LXRβ–/– mice. Animals were fed a standard rodent diet or a 2% cholesterol-enriched diet for 28 days. Livers were removed immediately after the animals were sacrificed.
(a and b) Total cholesterol content in hepatic lipid extracts and in serum. Animals were fed a standard rodent diet or 2% cholesterol-enriched diet for 28 days. Livers were removed immediately after the animals were sacrificed. Blood was obtained by cardiac puncture and serum was collected by centrifugation. Total cholesterol was determined enzymatically in hepatic lipid extracts and serum from ten animals per experimental group. All values are expressed as mean + SEM. P values are versus wild-type. The results from two independent analyses are shown in a and b. NS, not significant.
Analysis of serum cholesterol again showed that LXRβ–/– mice responded in the same way as wild-type controls after dietary cholesterol (LXRβ–/– vs. wild-type, not significant; Power >95%; Figure 3a). This finding was further confirmed by comparable plasma lipoprotein profiles in these two categories of animals (Figure 4). In LXRα–/– mice an increase in serum cholesterol was found in response to cholesterol feeding (Figure 3b), especially LDL, as seen upon analysis of lipoprotein profiles (Figure 4).
Serum lipoprotein profiles after separation by FPLC in LXRβ–/– (top) and LXRα–/– (bottom) mice. Animals were fed standard rodent diet (left) or challenged with 2% cholesterol (right) for 28 days. Ten microliters of pooled serum from each group was directly separated on a FPLC Superose 6B column. Cholesterol content was determined using the commercially available kit MPR 21442350 (Roche Molecular Biochemicals, Indianapolis, Indiana, USA) that was mixed online with the separated lipoproteins at a flow rate of 40 + 40 μl/min. The mixture was passed over a 37°C reaction coil and adsorbance was monitored every 20 seconds at 500 nm. The adsorbance profiles from two independent analyses are shown in the upper and lower panels.
Dietary cholesterol did not increase serum alanine amino transferase (ALAT) activity in LXRβ–/– mice. LXRα–/– mice showed a distinct increase in serum ALAT activity after cholesterol feeding (Table 1).
Serum analysis in LXR-deficient mice
Recently, alternative pathways for the conversion of oxysterols into bile acids have been described, starting with hydroxylation in the 27 position of cholesterol in the liver and extrahepatic organs (34–36) or hydroxylation of cholesterol in the 24 position occurring in the brain (36, 37). Since 24-hydroxycholesterol has been demonstrated to be a potent ligand for both LXRs in vitro (6, 7), we wanted to investigate if disruption of the LXR genes would affect this pathway. Therefore, we determined the serum levels of 24- and 27-hydroxycholesterol in pooled samples from both LXR-mutant mouse lines. In LXRβ–/– mice, 24- and 27-hydroxycholesterol levels differed from those of wild-type controls. Conversely, an increased serum concentration of 24- and particularly 27-hydroxycholesterol was found in response to cholesterol feeding in LXRα–/– mice (Figure 5).
Measurement of 24-hydroxycholesterol (24-OH-Chol) (a) and of 27-hydroxycholesterol (27-OH-Chol) (b) in serum of LXRα–/– or LXRβ–/– mice and their respective wild-type controls. All values were obtained from pooled serum of the same animals as in Figure 3.
Analysis of mRNA expression of genes involved in the regulation of cholesterol, fatty acid, and bile acid metabolism was performed to identify alterations in both LXR mutants. Northern blot hybridizations were done with mRNA extracted from pooled livers of female mice. Signals were quantitated and related to GAPDH mRNA expression.
In LXRβ–/– mice on a standard diet, an increase in 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (1.4-fold), FPP synthase (1.4-fold), and squalene synthase (1.6-fold) was observed as compared with wild-type controls (Figure 6). Differences in hepatic expression of HMG-CoA reductase, FPP synthase, and squalene synthase mRNA were not observed in cholesterol-fed LXRβ mutants and LXRα mutants when fed with both diets.
Expression of various lipid-regulating genes and of genes coding for enzymes in the neutral and acidic bile acid pathway in LXRβ–/– mice (a), LXRα–/– mice (b), and corresponding wild-type controls assayed by Northern blot analysis. Used were 2 or 5 μg/lane poly A+-enriched RNA, extracted from a pool of livers from five female mice fed as indicated. Filters were hybridized with [α-32P] dCTP-labeled cDNA probes, generated as described in Methods. Signals were standardized to the corresponding GAPDH expression. Data show the fold change compared with the respective wild-type control group fed the same diet. The results are representative of two or more independent determinations.
We next addressed the question of whether the expression of the lipogenic genes SCD-1, FAS, acetyl CoA carboxylase, and the transcriptional regulator ADD-1/SREBP-1 was altered by inactivation of the LXRβ gene. Northern blot analysis showed that the mRNA levels of these genes were not altered in animals on standard diet. After cholesterol feeding, a 1.5-fold increased mRNA level of acetyl CoA carboxylase was observed in LXRβ–/– mice (Figure 6). Disruption of the LXRα gene resulted in a downregulation of SCD-1, while expression of FAS, acetyl CoA carboxylase, and ADD-1/SREBP-1 remained unchanged. Cholesterol feeding reduced SREBP-1 and FAS mRNA levels in LXRα–/– mice compared with their wild-types (Figure 6).
In addition, we determined the hepatic mRNA expression of some selected genes important in the neutral and acidic biosyntheses of bile acids: Cyp7a, Cyp7b, Cyp8b1, and Cyp27. LXRβ-mutant mice fed either with standard chow or 2% cholesterol showed no differences in the expression levels of these genes (Figure 6). In LXRα–/– mice on either diet, the expression levels of Cyp7b, Cyp8b1, and Cyp27 were similar to those of wild-type animals. In contrast to what was observed in LXRβ–/– mice, cholesterol feeding reduced the mRNA level of Cyp7a in the liver of LXRα–/– mice (Figure 6).
In this report we compare the function of the LXR receptors in hepatic cholesterol metabolism in vivo. We have focused particularly on responses after cholesterol feeding. We generated two mutant mouse lines bearing targeted disruptions of LXRα or LXRβ. Similar to what was described in a previous report on LXRα-deficient mice (10), we found LXRβ–/– mice to be fertile and show no apparent abnormal phenotypes. LXRβ–/– mice maintained a normal response following dietary cholesterol. Both on standard diet and on cholesterol-enriched diet, livers of LXRβ–/– mice preserved a normal appearance. After cholesterol challenge, hepatic cholesterol concentrations increased in a way similar to their wild-type controls. This was associated with normal serum cholesterol levels and with normal lipoprotein profiles. These results are therefore in striking contrast to the data obtained from cholesterol-fed LXRα–/– mice. As reported previously (10) and as confirmed in this study, a dramatic increase in hepatic cholesterol content and a pronounced elevation of serum cholesterol levels, especially in the LDL fraction, were indeed observed in this animal model.
In females of both wild-type and LXRβ–/– mice, a trend toward a larger increase in hepatic cholesterol levels than in corresponding male mice was observed in response to cholesterol feeding. This is in agreement with data from Turley et al. (38), showing sex-related differences in response to cholesterol feeding in 129/Sv and C57BL/6 inbred mice. A larger bile acid pool size and a consequently increased intestinal cholesterol absorption have been proposed to be responsible for a more pronounced increase in hepatic cholesterol concentration in female mice on cholesterol feeding (38). Such a sex-related difference in hepatic cholesterol was not observed in LXRα–/– mouse mutants following cholesterol feeding, probably since deletion of the LXRα gene resulted in such extensive metabolic perturbations that possible sex-related differences would not be apparent.
High levels of serum 27-hydroxycholesterol following cholesterol feeding in LXRα–/– mice represent another marked difference between LXRα–/– and LXRβ–/– mice. The observation of unchanged mRNA levels for Cyp27 and Cyp8b1 indicate that a new dynamic equilibrium between cholesterol as substrate and its oxysterol products may have been reached in LXRα–/– mice. The level of 27-hydroxycholesterol in the circulation is dependent upon three factors: the activity of the synthesizing enzyme sterol 27-hydroxylase, the activity of the metabolizing enzyme oxysterol 7α-hydroxylase, and the level of cholesterol. The level of cholesterol is of importance since 27-hydroxycholesterol is transported in the same lipoproteins as cholesterol, and there is a relatively constant ratio between cholesterol and 27-hydroxycholesterol levels in humans (39). In LXRα–/– mice fed high cholesterol, not only the absolute but also the cholesterol-related levels of 27-hydroxycholesterol were elevated. Thus the observed effect cannot be a dilution effect only. Since our results excluded a downregulation of hepatic Cyp7b expression, decreased metabolism of 27-hydroxycholesterol cannot be the explanation for the increase. An increased formation of 27-hydroxycholesterol is thus a probable explanation for the increased level of these oxysterols in the circulation. Furthermore, since the LXRα-deficient mice did not have an increased expression of the Cyp27 gene, an increased formation of 27-hydroxycholesterol likely is due to an increased enzyme activity, possibly resulting from the higher availability of the substrate, cholesterol. It must be assumed that the mitochondrial sterol 27-hydroxylase is not normally saturated with its substrate. The situation may be similar for oxysterol 24(S) hydroxycholesterol.
To explain the abnormal response of LXRα–/– mice to cholesterol feeding, it has been proposed that the normal increase in bile acid synthesis — secondary to an upregulation of Cyp7a expression — is greatly reduced in these animals (10). In cholesterol-fed LXRβ–/– mice, the mRNA levels of Cyp7a, Cyp7b, and Cyp8b1, as well as the enzymatic activity of CYP7A (data not shown), were comparable to those of wild-type controls, indicating a normal hepatic bile acid metabolism. Our results are, however, in disagreement with published data showing an increase of Cyp7a gene expression after cholesterol feeding (40), since this was not observed in either wild-type controls or in LXRβ–/– mice. Furthermore, in cholesterol-fed LXRα–/– mice, Cyp7a mRNA expression was reduced in contrast to was what found by Peet et al. (10). A possible explanation for this discrepancy could be that animals were fasted 5 hours before sacrifice, since fasting reduces bile acid synthesis in rodents (41, 42).
The reduced Cyp7a mRNA expression in LXRα–/– mice when challenged with dietary cholesterol was not accompanied by a decreased Cyp27 mRNA expression, although a coordinated regulation of Cyp27 and Cyp7a genes by bile acids has been observed previously both in vitro and in vivo in the rat (43, 44). Recently, it has been demonstrated that bile acids can signal via the nuclear receptor FXR (45–47), and that this receptor is responsible for at least part of the negative feedback regulation of CYP7A by bile acids (47). If bile acids control the expression of Cyp27 through binding to FXR is still unknown. Nevertheless, an interesting possibility is that bile acids might regulate the expression of both Cyp27 and Cyp7a through FXR, whereas cholesterol would only control the expression of Cyp7a (via LXRα) but not of Cyp27. This notion is also supported by the observations on the dietary regulation of CYP7A and CYP27 in normal mouse liver (48).
Compared with their wild-type controls, LXRβ–/– mice on a normal diet show an increase in the mRNA levels of HMG-CoA reductase, FPP synthase, and squalene synthase, three genes encoding enzymes involved in cholesterol biosynthesis. These increases occurred despite normal expression of the genes for SCAP and the SREBPs, proteins known to regulate the transcription of the former three genes (49, 50). However, whether fed a standard or cholesterol-enriched diet, LXRβ-deficient male mice did not show an altered enzymatic activity of hepatic HMG-CoA reductase (data not shown).
In our study we observed variations in serum and hepatic cholesterol levels between the two wild-type control groups. The knockout mice and their corresponding wild-types were descendants from different progenitors, both having mixed C57BL/6-129/Sv backgrounds. Variations of the genetic background might influence the expression of genes involved in the regulation of cholesterol metabolism. Differences in cholesterol concentrations in hepatic tissue and plasma have been shown previously among inbred mouse strains, especially between C57BL/6 and 129/Sv mice (51). The partial recombinant inbred breeding as used in our study may accentuate these strain-specific differences. A previous study from Jolley et al. (52) demonstrated that cholesterol absorption differs among mice having pure C57BL/6 and 129/Sv background (52), possibly leading to a variability in cholesterol absorption in mice with mixed C57BL/6 and 129/Sv background. However, both C57BL/6 and 129/Sv mice respond to dietary cholesterol with increased plasma and hepatic cholesterol levels as well as increased bile acid synthesis, albeit with different magnitude (52). Furthermore, it should be noted that our data are generated in two independent experiments, performed at different times. Cholesterol homeostasis is regulated in a complex way. Therefore, the identification of factors responsible for variations in plasma and hepatic cholesterol levels observed in different experiments is very difficult, unless appropriate studies are performed. In an additional control experiment in which animals were sacrificed and serum determinations carried out at the same time, no differences in serum cholesterol of LXRα+/+ and LXRβ+/+ mice were observed (data not shown). Thus, the maintained resistance to cholesterol feeding in LXRβ–/– mice is less likely caused by the variability of the mixed genetic background of the progenitor, but is presumably due to the presence of an intact LXRα gene in these mice.
LXRβ, similar to LXRα, appears to be involved in the regulation of cholesterol metabolism in peripheral tissues, since both LXRs seem to have a critical role in the regulation of ABCA1 and ABCG1 (18–22). Preliminary data on cholesterol-challenged double-LXRα/LXRβ-mutant mice obtained by mating of mice with either the LXRα+/–/LXRβ–/– or LXRα–/–/LXRβ–/– genotype, revealed no obvious additive or synergistic effect compared with LXRα–/– alone (data not shown). This further supports the idea of no major overlap function of the LXRs in the regulation of hepatic cholesterol metabolism. Since appropriate control mice were not available, additional experiments should be performed to confirm these findings.
In conclusion, our data indicate that LXRβ, compared to LXRα, does not have the same important function in regulating the hepatic response to cholesterol challenge in mice, albeit subtle effects on the expression of enzymes involved in lipid metabolism were observed. Our findings are particularly interesting since both LXRs have a high degree of amino acid identity (17), similar ligand-binding affinity to endogenous oxysterols (6, 7), and redundant functions in macrophages and enterocytes (19, 21).
We thank Helena Sundén and Annika Samuelsson for ES-cell culture, and José Inzunza for blastocyst injections and implantations; Ralf Kühn and Klaus Rajewsky (Cologne, Germany) for the generous gift of pGEM30, pL2neo, and pIC-Cre vectors; Stefan Alexson and Christine Sadek for Northern probes; Anne Plück and Kristina Vintersten (Transgenic Core Facility, EMBL, Heidelberg, Germany) for delivery and implantation of cryo-preserved deleter mouse embryos, and Christine Sadek and Thomas Günther for critical reading of the manuscript. This work was funded by the Swedish Medical Research Council (13X-2819, 03X-3141, and 03X-7137), the Tore Nilson, Ruth and Bertil Julin, Fredrik and Ingrid Thuring, Ax:son Johnson, Swedish Heart-Lung foundations, the Foundation of Strategic Research, and KaroBio AB (Sweden).
S. Alberti and D. Feltkamp’s present address is: University of Tübingen, Institute for Cell Biology, Department of Molecular Biology, Tübingen, Germany.
Gordon, T, Kannel, WB, Castelli, WP, Dawber, TR. Lipoproteins, cardiovascular disease, and death. The Framingham study. Arch Intern Med 1981. 141:1128-1131.
Keys, A. Coronary heart disease, serum cholesterol, and the diet. Acta Med Scand 1980. 207:153-160.
Angelin, B. 1994 Mack-Forster Award Lecture. Review. Studies on the regulation of hepatic cholesterol metabolism in humans. Eur J Clin Invest 1995. 25:215-224.
Goldstein, JL, Brown, MS. Regulation of the mevalonate pathway. Nature 1990. 343:425-430.
Accad, M, Farese (Jr), RV. Cholesterol homeostasis: a role for oxysterols. Curr Biol 1998. 8:R601-R604.
Lehmann, JM, et al. Activation of the nuclear receptor LXR by oxysterols defines a new hormone response pathway. J Biol Chem 1997. 272:3137-3140.
Janowski, BA, Willy, PJ, Devi, TR, Falck, JR, Mangelsdorf, DJ. An oxysterol signalling pathway mediated by the nuclear receptor LXR alpha. Nature 1996. 383:728-731.
Forman, BM, Ruan, B, Chen, J, Schroepfer, J, Evans, RM. The orphan nuclear receptor LXRα is positively and negatively regulated by distinct products of mevalonate metabolism. Proc Natl Acad Sci USA 1997. 94:105888-105893.
A unified nomenclature system for the Nuclear Receptor Superfamily Cell 1999. 97:161-163.
Peet, DJ, et al. Cholesterol and bile acid metabolism are impaired in mice lacking the nuclear oxysterol receptor LXR alpha. Cell 1998. 93:693-704.
Shinar, DM, et al. NER, a new member of the gene family encoding the human steroid hormone nuclear receptor. Gene 1994. 147:273-276.
Seol, W, Choi, HS, Moore, DD. Isolation of proteins that interact specifically with the retinoid X receptor: two novel orphan receptors. Mol Endocrinol 1995. 9:72-85.
Song, C, Kokontis, JM, Hiipakka, RA, Liao, S. Ubiquitous receptor: a receptor that modulates gene activation by retinoic acid and thyroid hormone receptors. Proc Natl Acad Sci USA 1994. 91:10809-10813.
Teboul, M, et al. OR-1, a member of the nuclear receptor superfamily that interacts with the 9-cis-retinoic acid receptor. Proc Natl Acad Sci USA 1995. 92:2096-2100.
Willy, PJ, et al. LXR, a nuclear receptor that defines a distinct retinoid response pathway. Genes Dev 1995. 9:1033-1045.
Apfel, R, et al. A novel orphan receptor specific for a subset of thyroid hormone-responsive elements and its interaction with the retinoid/thyroid hormone receptor subfamily. Mol Cell Biol 1994. 14:7025-7035.
Alberti, S, Steffensen, KR, Gustafsson, J-Å. Structural characterization of the mouse nuclear oxysterol receptor genes LXRα and LXRβ. Gene 2000. 243:93-103.
Schwartz, K, Lawn, RM, Wade, DP. ABC1 gene expression and ApoA-I-mediated cholesterol efflux are regulated by LXR. Biochem Biophys Res Commun 2000. 274:794-802.
Venkateswaran, A, et al. Human white/murine ABC8 mRNA levels are highly induced in lipid-loaded macrophages. A transcriptional role for specific oxysterols. J Biol Chem 2000. 275:14700-14707.
Klucken, J, et al. ABCG1 (ABC8), the human homolog of the Drosophila white gene, is a regulator of macrophage cholesterol and phospholipid transport. Proc Natl Acad Sci USA 2000. 97:817-822.
Repa, JJ, et al. Regulation of absorption and ABC1-mediated efflux of cholesterol by RXR heterodimers. Science 2000. 289:1524-1529.
Costet, P, Luo, Y, Wang, N, Tall, AR. Sterol-dependent transactivation of the ABC1 promoter by the liver X Receptor/Retinoid X receptor. J Biol Chem 2000. 275:28240-28245.
Gu, H, Zou, YR, Rajewsky, K. Independent control of immunoglobulin switch recombination at individual switch regions evidenced through Cre-loxP-mediated gene targeting. Cell 1993. 73:1155-1164.
Gu, H, Marth, JD, Orban, PC, Mossmann, H, Rajewsky, K. Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. Science 1994. 265:103-106.
Mansour, SL, Thomas, KR, Capecchi, MR. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 1988. 336:348-352.
Nagy, A, Rossant, J, Nagy, R, Abramow-Newerly, W, Roder, JC. Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc Natl Acad Sci USA 1993. 90:8424-8428.
Schwenk, F, Baron, U, Rajewsky, K. A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. Nucleic Acids Res 1995. 23:5080-5081.
Folch, J, Lees, M, Stanley, GHS. A simple method for the isolation and purification of total lipids from animals tissues. J Biol Chem 1957. 226:497-509.
Dzeletovic, S, Breuer, O, Lund, E, Diczfalusy, U. Determination of cholesterol oxidation products in human plasma by isotope dilution-mass spectrometry. Anal Biochem 1995. 225:73-80.
Bergmeyer, HU, Horder, M, Rej, R. IFCC expert panel on enzymes. Part 2. J Clin Chem Clin Biochem 1986. 24:497-510.
Shimano, H, et al. Overproduction of cholesterol and fatty acids causes massive liver enlargement in transgenic mice expressing truncated SREBP-1a. J Clin Invest 1996. 98:1575-1584.
Marchuk, D, Drumm, M, Saulino, A, Collins, FS. Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Res 1991. 19:1154.
Snedecor, G.W., and Cochran, W.G. 1980. Statistical methods. Seventh edition. The Iowa State University Press. Ames, Iowa, USA. 507 pp.
Stravitz, RT, et al. Regulation of sterol 27-hydroxylase and an alternative pathway of bile acid biosynthesis in primary cultures of rat hepatocytes. J Steroid Biochem Mol Biol 1996. 57:337-347.
Axelson, M, Sjövall, J. Potential bile acid precursors in plasma: possible indicators of biosynthetic pathways to cholic and chenodeoxycholic acids in man. J Steroid Biochem 1990. 36:631-640.
Björkhem, I, Diczfalusy, U, Lütjohann, D. Removal of cholesterol from extrahepatic sources by oxidative mechanisms. Curr Opin Lipidol 1999. 10:161-165.
Lütjohann, D, et al. Cholesterol homeostasis in human brain: evidence for an age-dependent flux of 24S-hydroxycholesterol from the brain into the circulation. Proc Natl Acad Sci USA 1996. 93:9799-9804.
Turley, SD, Schwarz, M, Spady, DK, Dietschy, JM. Gender-related differences in bile acid and sterol metabolism in outbred CD-1 mice fed low- and high-cholesterol diets. Hepatology 1998. 28:1088-1094.
Harik-Khan, R, Holmes, RP. Estimation of 26-hydroxycholesterol in serum by high-performance liquid chromatography and its measurement in patients with atherosclerosis. J Steroid Biochem 1990. 36:351-355.
Rudling, M. Hepatic mRNA levels for the LDL receptor and HMG-CoA reductase show coordinate regulation in vivo. J Lipid Res 1992. 33:493-501.
Björkhem, I. 1985. Mechanism of bile acid synthesis in mammalian liver. In New comprehensive biochemistry. Volume 12. Elsevier Science Publishing Co. Amsterdam, The Netherlands. 231–278.
Myant, NB, Mitropoulos, KA. Cholesterol 7 alpha-hydroxylase. J Lipid Res 1977. 18:135-153.
Twisk, J, et al. Heterogeneous expression of cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase genes in the rat liver lobulus. J Clin Invest 1995. 95:1235-1243.
Twisk, J, et al. Structural aspects of bile acids involved in the regulation of cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase. Eur J Biochem 1995. 228:596-604.
Wang, H, Chen, J, Hollister, K, Sowers, LC, Forman, BM. Endogenous bile acids are ligands for the nuclear receptor FXR/BAR. Mol Cell 1999. 3:543-553.
Parks, DJ, et al. Bile acids: natural ligands for an orphan nuclear receptor. Science 1999. 284:1365-1368.
Makishima, M, et al. Identification of a nuclear receptor for bile acids. Science 1999. 284:1362-1365.
Schwarz, M, Lund, EG, Russell, DW. Two 7 alpha-hydroxylase enzymes in bile acid biosynthesis. Curr Opin Lipidol 1998. 9:113-118.
Brown, MS, Goldstein, JL. The SREBP pathway: regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor. Cell 1997. 89:331-340.
Hua, X, Nohturfft, A, Goldstein, JL, Brown, MS. Sterol resistance in CHO cells traced to point mutation in SREBP cleavage-activating protein. Cell 1996. 87:415-426.
Kirk, EA, et al. Hyper- and hypo-responsiveness to dietary fat and cholesterol among inbred mice: searching for level and variability genes. J Lipid Res 1995. 36:1522-1532.
Jolley, CD, Dietschy, JM, Turley, SD. Genetic differences in cholesterol absorption in 129/Sv and C57BL/6 mice: effect on cholesterol responsiveness. Am J Physiol 1999. 276:G1117-G1124.

domain: biology


Chapman gets year back
Of the Alabama freshmen who saw action only at the begining of last season, Josh Chapman's hopes of a medical redshirt looked as slim as any.
After all, the Crimson Tide noseguard played in three games - two more than some others, and just barely within NCAA guidelines for the provision that allows freshmen who are injured to obtain a year of eligibility back despite seeing action.
According to the just-released UA media guide, Chapman (6-foot-1, 305 pounds) will return as a freshman in 2008. It is unclear which other freshmen UA applied for medical redshirt status on, but those who played only early in the season include Chapman, quarterback Nick Fanuzzi, defensive end Alfred McCullough, wide receiver Darius Hanks and offensive lineman William Vlachos.
According to the fall roster, Hanks and McCullough will be sophomores this fall, while Vlachos and Fanuzzi will be freshmen. All four played in a season-opening win against Western Carolina only.
Chapman played against Western Carolina, Vanderbilt and Arkansas as the Crimson Tide opened the 2007 season at 3-0.
FRESHMAN JERSEYS - A look at the jersey assignments listed for UA freshmen in the media guide: Mark Barron (4), Undra Billingsley (94), John Michael Boswell (97), Terrence Cody (62), Marcel Dareus (57), Robby Green (23), Glenn Harbin (54), Jerrell Harris (5), Don'ta Hightower (56), Mark Ingram (22), Star Jackson (2), Barrett Jones (75), Julio Jones (8), Chris Jordan (36), Alonzo Lawrence (15), Robert Lester (37), Tyler Love (72), Ivan Matchett (31), Wesley Neighbors (46), Jermaine Preyear (44), Burton Scott (1), Damion Square (65), Courtney Upshaw (41), Michael Williams (89).


text:
Best Spring 2014 Theater Productions In BaltimoreBaltimore theaters are putting on plays for your entertainment with well-known productions, both old and new, for theater aficionados and newcomers alike.
CENTERSTAGE Among U.S. Theaters Offering Discounts To Military FamiliesA program to offer free and discounted theater tickets, classes and other services for veterans and military families is expanding to 85 theaters in 33 states.
On Time: Sunday, May 1, 2011Kai Jackson spoke with Irene Lewis and Mark Feiring.
Best Baltimore Concert Guides For KidsSome of the best Baltimore concert guides for kids are: the Baltimore Symphony Orchestra, CENTERSTAGE, the Lyric Opera House, the Hippodrome Theater and the Maryland Hall for the Creative Arts.

domain: entertainment


Complying with a directive from the White House, the Department of Labor on Monday released a proposed rule and interpretive bulletin to help guide states in developing state-run retirement plans that don’t run afoul of ERISA.
Noting the nearly 70 million workers without access to retirement plans at work, Labor Secretary Thomas Perez said in releasing the plan that employees’ inability to save through work is not only a “potential financial crisis for these individuals and their families, but a critical economic issue for the nation.”
Monday’s guidance, Perez said, “is another plank in the economic security platform that President Obama and this administration have been building to help create new savings options, ensure workers are getting sound retirement advice, and bolster bedrock programs such as Social Security.”
DOL filed Sept. 3 with the Office of Management and Budget its proposed rule designed to make it easier for states to offer their own retirement plans without running afoul of the Employee Retirement Income Security Act.
During the White House’s Conference on Aging, held in July, President Barack Obama directed Perez to publish a proposed rule to “provide a clear path forward for the states to create retirement savings programs” by year-end.
Specific to the proposal is a new safe harbor from ERISA for state-sponsored IRAs that conform to certain provisions. The proposal would adopt a standard stating that state-sponsored payroll deduction IRA programs must be “voluntary” for workers, rather than “completely voluntary” as defined in a 1975 rule.
“This will allow for automatic enrollment of employees in such programs so long as they are given the ability to opt out, and employers are minimally involved,” DOL states.
For instance, “employers would make the automatic deductions from employee paychecks, but the employees and states would retain control of the program and IRA accounts,” DOL says. “Employers could not prevent workers from declining to participate in the program.”
Comments on the proposed rule are due by Jan. 19.
DOL also published an interpretive bulletin regard creating state-based ERISA-compliant 401(k) plans that are open to businesses and workers. In addition to payroll deduction IRAs and state-based 401(k)s, the bulletin provides examples of how to create state retirement savings programs that may avoid being preempted under ERISA.
Illinois Treasurer Michael Frerichs said that nearly 1.2 million workers are poised to benefit from Illinois’ state-run plan, the Secure Choice Retirement Savings Program. “With these new federal rules, my administration can move forward with helping some of our state’s most vulnerable workers,” Frerichs said.


text:
Beyond Body Weight: Design and Validation of Psycho-Behavioural Living and Eating for Health Segments (LEHS) Profiles for Social Marketing
Authors: Linda Brennan, Shinyi Chin, Annika Molenaar, Amy M. Barklamb, Megan SC Lim, Mike Reid, Helen Truby, Eva L. Jenkins and Tracy A. McCaffrey.
Background: Obesity, sedentary behaviour, and poor dietary habits amongst young adults are growing concerns, with this age group being in a worse state of health and nutrition than adolescents and adults. This paper presents the procedures for establishing a new instrument for defining behaviours in relation to healthy lifestyle and food choices amongst young adults (Living and Eating for Health Segments: LEHS).
Aim: The aim of this paper is to outline the instrument design protocol for external validation and to permit replication in other studies.
Methods: The instrument design process used a multi-step social marketing instrument design method. This approach has previously been used in designing valid and reliable measures in marketing and consumer research, including social marketing.
Results: The protocol established six psycho-behavioural LEHS profiles for young adults. These profiles are: Lifestyle Mavens (15.4%), Aspirational Healthy Eaters (27.5%), Balanced-all Rounders (21.4%), Health Conscious (21.1%), Contemplating Another Day (11.2%), and Blissfully Unconcerned (3.4%).
Conclusions: Each of these profiles provided insights into psycho-behavioural characteristics that can be used in designing apposite social media social marketing campaigns.
Keywords: Social marketing; social media; healthy eating; young adults; obesity prevention; instrument development.

domain: health


The Power Networks Demonstration Centre - Temperature and Humidity Monitoring in Electricity Substations
Published: 18 September, 2020
Case study supplied by Lascar Electronics
The University of Strathclyde Power Networks Demonstration Centre (PNDC) is a research, demonstration and deployment centre for emerging electrical power system technologies and accelerating smart grid implementations.
Founded by government, industrial and academic partners, the facility provides a realistic electrical network test system, and plays a leading role in the development of a smart electricity grid and the acceleration of low carbon technology.
To realise the transformation towards a smart grid, they have introduced a next generation communications platform which makes use of sensor technologies, and addresses the challenges of security, resilience and efficiency in electricity supply. Their work includes modelling of electricity grids and naval ship power systems, as well as testing new technology with a view to it reaching commercial deployment.
As part of this brief, the way in which temperature and humidity is measured inside a primary substation was looked at.


text:
LETTER — Dog owners shouldn't be allowed to leave their pets in hot cars
On July 31 I attended the Ottawa County Fair. On our way in we passed a vehicle with a husky inside with the windows barely cracked, and it was a hot day. We told security and they made one announcement, while drag racing was going on. After two hours we left and sure enough the dog was still in the car panting very badly. We informed the police on our way out but my guess is nothing was done.
I believe the vehicle should not have been allowed onto the grounds with the dog in the car. There are 14 states where this is illegal and punishable by fines and jail time. Michigan is not one of them. We need to do something to protect these animals.

domain: law


Your activewear has a tough job. You expect your tank tops, running shorts, underwear, and hoodies to let you move and keep you comfortable during your workouts and throughout your day, no matter what pace you’re putting them through.
It’s a tough job, absorbing the sweat, oils, and dead skin that your body sheds when you’re working out. So you might think that you just accept the fact that your sportswear becomes a bit funky over time. But it doesn’t have to be that way. If you know how to wash your sportswear the right way, it will stay a lot fresher, a lot longer, plus your favorite sweatpants will last longer too.
When you put on a high-quality piece of activewear, like bike shorts or short sleeve t-shirts from a best selling brand like Willy California, you can tell right away that they’re built to perform. The cut is exact, the fabric light, and they have a certain degree of stretch you won’t find in a cotton crew neck tee shirt or a pair of denim jeans. These attributes help make activewear perform the way you want it to, but they also make washing and care more difficult. Let’s examine some of the differences more closely.
The paradox of quality activewear is that it’s durable enough to stand up to hard use, but delicate when it comes to cleaning and care. That’s a result of the very specialized fabrics that are used to make activewear. In particular, many pieces of athleisure like leggings and yoga pants use spandex or elastane to add support and stretchiness. Both of these materials can be damaged by laundry additives like fabric softeners or by excessive heat.
To achieve the light weight that we prize in our activewear, manufacturers need to employ thin and light fabrics that are moisture-wicking. Even when these fabrics are durable, they won’t hold up to repeated washings as well as clothing made from heavier cloth.
Tougher Cleaning Demands
Of course, one of the biggest problems you encounter in washing activewear is the degree of cleaning you’ll need to do. If you’re working up a sweat at the gym, imagine what effect your perspiration is having as it soaks into your clothes. Your sweat and oils from your skin create a perfect environment for the creation of odor-causing bacteria. If you don’t wash your activewear properly, these bacteria will flourish and create a sour, musty smell that is hard to get rid of.
There’s a right way to wash your workout gear, but it’s a good idea to look at some of the common mistakes that we often make when doing our laundry. Eliminating these mistakes is an important way to stay on the right foot going forward.
Don’t Mix Fabrics
You may be tempted to toss your activewear into the wash along with your gym towels and other items, like socks, sweatshirts, swimwear, lingerie, or jeans. That’s a big mistake. Heavy items like denim jeans and towels can damage the lighter fabrics used in your activewear, causing pilling. And clothes with stretch fabrics like jogger shorts are prone to trapping lint from cotton clothing when you put them all in the same load.
Don’t Use Too Much Detergent
Your clothes are dirty? Pour in a little extra detergent, right? Not so fast. Too much detergent is actually bad for your activewear wardrobe. Instead of making your clothes cleaner, all you’re doing is adding a layer of soap residue that traps odors. The same goes for fabric softeners — keep them far away from your activewear ensembles.
Avoid High Temperatures
Clothes with elastic fibers like high rise capri leggings, biker shorts, v-neck t-shirts, and strappy racerback sports bras do not handle heat well. When washing your activewear and loungewear, go with cold water, and tumble dry low. Or even better, air dry your clothing.
Ready to get started washing your activewear the right way? Just follow this guide and you won’t go wrong!
Turn Your Gear Inside Out Before You Start
Taking just a few moments to turn your clothing inside out before washing will make a big difference in how clean they become. The reason is simple. With athletic gear, the real action is against your skin. That’s where sweat, oils, and dead skin cells accumulate, so you want to expose this side to the benefits of your wash.
Use the Right (and the Right Amount) of Detergent
Don’t guess about how much detergent to use. Read the instructions and then measure out a bit less than the recommended amount. And consider investing in some detergent that’s made for activewear. You may spend a bit more, but the results will be better. Also pay close attention to color and designs on your sportswear like logos and graphic tees, camo, and tie-dye shirts to make sure that those features survive the wash.
Use the Right Settings on Your Washing Machine
You did read the label on your clothing, right? If you didn’t check the care label so you get the right water temperature. If you’re in doubt, go with a cold water wash that works well with everything from dri fit long sleeve t-shirts to sleeveless crop tops.
Skip the Dryer and Air Dry If Possible
Your clothes will last longer if you skip the dryer, plus there’s less opportunity to “bake in” odors. Invest in a folding drying rack if you don’t have a clothesline available.
With the right care, your workout clothes will last longer, look better, and smell fresher. It’s simple — if you care enough to invest your money in high-quality activewear why not invest your time in learning how to wash and care for it properly? Take care of your gear and you’ll always look good when you hit the gym. The rest is up to you.


text:
Market Report: To value AB Foods, you must look at the sum of its parts... the 'jewel' isPrimark
Thursday 07 November 2013
The City can be fickle, and after falling out of love with cheap clothes to bread maker Associated British Foods on Tuesday, yesterday the love affair was back on.
At least five analysts formed an orderly queue to praise the Primark to Kingsmill group, a day after it lost 2 per cent as investors worried its sugar unit is dragging on growth.
But AB Foods was 61p sweeter at 2,269p yesterday as scribblers focused on its long-term prospects – namely the international potential of Primark. The Bangladesh garment factory disaster hasn't put off shoppers, and its growth continues to be phenomenal. UBS said "the best way to value AB Foods is to look at the sum of its parts… the 'jewel' is Primark".
Another stock to benefit from a change of heart by analysts was security group G4S. Positive comments from JPMorgan's experts – helped it top the benchmark index, up 7.8p at 257.5p.
The wider market followed positive European sentiment ahead of today's European Central Bank update. But the FTSE 100 finished down 5.15 points at 6,741.69.
Blue-chip heavyweight Vodafone benefited from continued talk of a bid from the US telecoms giant AT&T, dialing up 2.35p to 230.6p, while a jump in monthly passenger numbers at Easyjet, up 11p at 1,214p, also helped British Airways owner IAG and Tui Travel rise 4.7p to 349.8p and 8.5p to 383.6p respectively.
Experian, however, dropped 81p to 1,182p as it said conditions were subdued in some emerging markets and revealed it had paid £528m for a US healthcare firm Passport Health Commnunications.
On the mid-tier table, the worst performer was gold miner Centamin, down 2.5p to 48.3p. News that shareholder Nyota Minerals has cut its stake weighed even as it reported gold production for the third-quarter was up 39 per cent year on year.
The pubs group JD Wetherspoon rose 7.5p to 720p as it reported a comparable sales rise of 3.7 per cent and said it will continue to open new pubs.
Hovis and Mr Kipling owner Premier Foods confirmed it has appointed Ondra Partners to look at options to sell its bread business and it gobbled up 4.5p to 144.5p.
- 1 VMAs 2015: Was Nicki Minaj and Miley Cyrus' awkward acceptance put-down real or staged?
- 2 If you're not already angry about the migrant crisis, here's a history lesson to remind you why you really should be
- 3 Rules on 5p plastic bags likely to lead to arguments at the check-out
- 4 I like Corbyn, but let's face it: we don't need another white man at the head of a political party
- 5 Orthorexia nervosa: How becoming obsessed with healthy eating can lead to malnutrition
VMAs 2015: Was Nicki Minaj and Miley Cyrus' awkward acceptance put-down real or staged?
Bank Holiday Monday opening times: Are Tesco, Asda and other supermarkets open today?
Rules on 5p plastic bags likely to lead to arguments at the check-out
The nine most warmongering countries in the world revealed
VMAs 2015: Kanye West runs for president, Nicki Minaj calls out Miley Cyrus and the list of winners in full
Climate change: 2015 will be the hottest year on record 'by a mile', experts say
'Women only' train carriages: Jeremy Corbyn unveils radical move to tackle public harassment
Black holes are a passage to another universe, says Stephen Hawking
Tony Blair attacks Jeremy Corbyn's 'Alice In Wonderland' politics
Theresa May says migrants should be banned from entering the UK unless they have jobs lined up
Iain Duncan Smith 'should resign over disability benefit death figures', says Jeremy Corbyn
iJobs Money & Business
£25000 - £30000 per annum: Recruitment Genius: From modest beginnings the comp...
£35000 - £40000 per annum: Recruitment Genius: From modest beginnings the comp...
£15000 - £65000 per annum: Recruitment Genius: This is an exciting opportunity...
£18000 - £20000 per annum: Recruitment Genius: This is a fantastic opportunity...

domain: news


text:
Read Gateway. Kroniki Heechów by Frederik Pohl Free Online
Book Title: Gateway. Kroniki Heechów|
The author of the book: Frederik Pohl
Format files: PDF
The size of the: 413 KB
City - Country: No data
Loaded: 2831 times
Reader ratings: 5.7
Date of issue: 2001
ISBN 13: 9788388431180
Read full description of the books:
I must confess, I liked this book enough to finish it but I am glad the series is finished. Robinette Broadhead is a dull, histrionic main character and this book contains waaaay too much lecturing and philosophizing. A lot of that is done through Pohl’s usual device in this series: the computer program known as Albert Einstein.
You would think that when Rob Broadhead died, we would be free of him as main character. Instead, he is uploaded into a computer environment and survives to remain the focus of the series. I do take issue with a stored intelligence being as moody, weepy, and “gloopy” as Pohl writes Rob—without the brain structures to support emotions, I just doubt that emotion would be able to run someone’s life so completely. But what do I know? We are so far away from being able to do this, that there isn’t any way of judging. And all this anxiety now seems to be pointless—Rob is basically immortal, still wealthy & influential, and has his lady love stored with him. His only worry should be maintaining the machinery that comprises his new environment and keeping up to technological changes.
And OMG, the astrophysics that gets booted around this novel! I never minded it from Arthur Clarke, who made it a part of the story without long info-dump interludes, but Pohl gives Albert way too much screen time. Although he keeps telling Rob that he’s “making it as simple as he can,” I get the sense that Pohl was enjoying giving little lectures and I really wish that his editor had been more brutal.
What I did enjoy was getting a better sense of the Heechee, although I have still never figured out where that name came from. Pohl introduces other life forms as well, the Sluggards and the Voodoo Pigs among them. I think he could have used some help with better names for his aliens and with making them more interesting and relevant to the story. Having said that, when we do finally discover life elsewhere in the universe, I wonder how much of it will be bacteria, algae, or coral-like? Not really something that we can interact with in a meaningful way. But this is fiction, and I persist in wanting interesting aliens (like in Brin’s Uplift universe).
What a contrast to the work of Arthur Clarke! Pohl’s work is sticky with emotion, while Clarke’s is cooly intellectual. Both Pohl and Clarke have humans dealing with vastly superior alien civilizations, but Pohl’s seems menacing while Clarke’s seem to be mildly interested (2001, Childhood’s End) to indifferent (Rendezvous with Rama). Clarke’s work left me longing to know more about these aliens, while Pohl’s left me dissatisfied that they had been so involved, and yet I knew next to nothing about them.
Book number 226 in my science fiction & fantasy reading project.
Download Gateway. Kroniki Heechów ERUB
Download Gateway. Kroniki Heechów DOC
Download Gateway. Kroniki Heechów TXT
Read information about the author
Reviews of the Gateway. Kroniki Heechów
Add a comment
Download EBOOK Gateway. Kroniki Heechów by Frederik Pohl Online free

domain: literature


text:
By Josh Speer / February 18th, 2013
Nintendo and Retro Studios have unveiled the following trailer for their upcoming title Donkey Kong Country Returns 3D, set to come out for the 3DS this summer. Looks like the graphics hold up pretty well on the portable version.
Video courtesy of Official Nintendo Magazine
3DSDonkey Kong Country Returns 3DNintendoRetro StudiosTrailer


text:
Just wanted to ask if the book lists all the names of the cemetaries in Fairfield? And does it give descriptions of where they are located?
As for the Mitchell's I would be interested in more info. I have found John benjamin Mitchell's(My 4th great grandfather) gravestone and jemima's. But for some reason Jemima was moved from Morey cemetary and buried in little Egypt. Does anyone know why or does the book say??
I was told to check Gilbert Harry Doane's books out. Guess I'm going to go to the library on my next day off.

domain: library


To celebrate, Bandai releases 2 new editions of the game.
Tekken 7, a fighting game developed and published by Bandai Namco Entertainment and released in March 2015, has sold over 8 million copies, reports the developer. This is a huge success and we congratulate the game and its creators on achieving this milestone.
To celebrate, Bandai releases 2 new editions of the game – Originals and Definitive. A new trailer published by the studio on its official YouTube channel highlights them both and sheds some light on the Definitive Edition, which comes with all the DLC content the game has ever had.
According to the official description on the video, Tekken 7 Originals Edition contains the main game, 12 Character Pack, and Frame Date Display, the Definitive Edition, on the other hand, has the main game, Eliza Bonus, Season Pass 1-4, plus bonuses.


text:
*From Troy Media Relations*
BOWLING GREEN, Ky. – Western Kentucky scored a run in the fifth and sixth innings to rally past Troy 5-4 Saturday afternoon at Nick Denes Field and even the three-game Sun Belt Conference series at a game each.
Ali Knowles gave Troy a 4-3 lead in the third with a two-out single to left after back-to-back two-out base hits by Matthew Howard and Garrett Pitts.
Trevor Lowe led off the WKU (19-16, 9-5 Sun Belt) half of the fifth with a triple just off the glove of Danny Collins in right field and Steve Hodgins followed with a single up the middle to knot the game at 4-all.
A leadoff walk to Devin Kelly in the sixth proved to be the difference after he went first to third on a sac bunt and scored on a single by Jordan Cessna.
Collins reached in both the seventh and ninth innings, but was not be able to come around and score the tying run. The junior slugger put the Trojans (24-10, 8-6) on the board with a three-run homer in the first, it was Collins’ eighth home run of the season as was the start of a 3-for-5 day.
The Hilltoppers matched the Trojans’ three-run first with one of their own. Ryan Huck put WKU on the board with a single to left after the first two batters of the inning reached on seeing-eye singles. Following a pair of outs, Blake Crabtree tied the game with a two-run single to center field.
WKU stretched starter Andrew Edwards to the limit as the junior threw 132 pitches over eight innings of work. Edwards (4-1) allowed all four runs, three earned, on eight hits; he walked three – all in the first inning – and struck out six. Austin Clay recorded the final out of the game for his second save of the year.
Nate Hill (1-2) took the hard-luck loss for Troy despite pitching the Trojans out of a sticky situation in the fifth. The southpaw inherited runners on first and third with just one out. Hill got a groundout and a fly out around an intentional walk to Huck to end the threat.
Ryan Sorce gave the Trojans 3.1 quality innings out of the bullpen. He scatter four hits, allowed one run and struck out three after entering the game in the second inning.
The teams will play the rubber game of the series Sunday afternoon at 1 p.m.


Debian Security Advisory
DSA-2806-1 nbd -- privilege escalation
- Date Reported:
- 29 Nov 2013
- Affected Packages:
- Security database references:
- In Mitre's CVE dictionary: CVE-2013-6410.
- More information:
It was discovered that nbd-server, the server for the Network Block Device protocol, did incorrect parsing of the access control lists, allowing access to any hosts with an IP address sharing a prefix with an allowed address.
For the oldstable distribution (squeeze), this problem has been fixed in version 1:2.9.16-8+squeeze1.
For the stable distribution (wheezy), this problem has been fixed in version 1:3.2-4~deb7u4.
For the unstable distribution (sid), this problem will be fixed soon.
We recommend that you upgrade your nbd packages.


text:
| Uttar Pradesh|
Majhola is a class III town and a nagar panchayat in Pilibhit district in the Indian state of Uttar Pradesh. This town is located 25 km away from district headquarters on state highway 29 connecting Pilibhit to Tanakpur town in Uttaranchal. It is well connected by metre gauge railway line to District headquarters. Geographically, town is located adjacent to Uttaranchal border in terai region. Terai region is famous for its fertile land and industrious farmers. Agriculture is the mainstay of the town economy. It has one sugar factory in cooperative sector, one distillery and few flour mills. Both sugar factory and distillery are now defunct. Demographic profile of town consists of people from different social groups. Dominant among them are farmers which are mainly Sikhs and middle caste group migrants from eastern Uttar Pradesh. Others are business class upper caste people from western Uttar Pradesh who dominates the town economy.Town has two Intermediate college and one convent school. There are many place of worships belonging to different religions. The place had very vibrant sport culture which slowly diminished due to exodus of youth out of town for employment.
As of 2001 India census, Majhola had a population of 21,824. Males constitute 52.45% of the population and females 47.55%. Nyoria Husainpur has an average literacy rate of 29%, lower than the national average of 59.5%: male literacy is 37%, and female literacy is 20%. In Majhola, 16% of the population is under 6 years of age.

domain: geography


IBE, an Internet Booking Engine solution framework developed by Hexaware, is an outcome of our airline domain expertise. IBE helps the travel and tourism industry support online reservation and related services for consumers. Travel services providers can enhance their customer reach through an extended distribution system as well as offer a richer customer experience while leveraging state-of-the-art technologies to reduce operational costs.
IBE implements comprehensive Service Oriented Architecture (SOA) and can seamlessly fit into the airline’s existing infrastructure. Business / IT agility is enabled by externalising the process flows and providing a loosely coupled process design with flexibility to integrate with third party databases such as GDS or non-GDS vendors and reservation host systems. Our solution can be customised to fit into current airline requirements or future changing market needs.
IBE provides an intuitive interface for the customer, making the booking process simple and quick. It uses a relational database to provide flexibility to cache the airline schedule and inventory and fare data for minimising communication with the GDS/reservation host system. The result? A dramatic increase in the speed of the booking engine and significantly lower costs.
- Airline Schedules: Provides customer the ability to search flight schedules
- Availability by Fare: Customer can select fares and view corresponding applicable flights
- Profile administration: Provides the capability to use customer profiles (corporate and individual) for improved booking experience
- Airline Seat Maps: Offers colorful airline seat maps that help customers choose seating in advance and provides a better perception of the in-flight service experience
- Intelligent Caching: IBE’s local database provides flexibility to cache the airline schedule, inventory and fares data to minimise communications with the GDS/reservation host system and keep transactional costs to a minimum
- Payment Processing: Connection to third party payment gateways (credit card) and inventory is sold only if payment authorisation occurs
- Special Service Request (SSR): Configuration of special services and processing
- Localisation: Multi-lingual
- Host System: Operates with any legacy host reservation system
- Customisable look and feel
- A complete booking engine
- Frequently accessed information is cached to minimise access to GDS
- Pre-filled preferences for existing customer profiles reducing manual input
- Business / IT agility enabled by Service Oriented Architecture (SOA) implementation
- Business rules externalised by airline configurable parameters leading to scalability
- Seamless integration with external interfaces
- Easy Administration
Contact us to learn more about Hexaware’s Industry Point Solutions and our deck of success stories


text:
Adam Pekor is an attorney in Labor and Employment Practice Group in the firm's New York office.
Areas of Practice
Adam counsels both for-profit and not-for-profit clients on a wide range of labor and employment matters, including employee hiring and disciplinary procedures, disability accommodation issues, restrictive covenants, worker classification issues, leaves of absence, wage and hour compliance, and employee severance and termination procedures. Adam has significant experience drafting all types of employment agreements and policies, including employee handbooks, confidentiality and non-disclosure agreements, severance agreements, sexual harassment policies, and email and internet usage policies. Adam also conducts discrimination and harassment prevention trainings as well as investigations into harassment allegations and other employment matters.
In addition, Adam regularly represents clients in a variety of labor and employment disputes before federal and state courts and administrative agencies, as well as in arbitration proceedings.
New York Rising Star, Super Lawyers, 2016-2019
Labor & Employment Law Blog Posts
- "Second Circuit Reiterates: Bigger Hurdles For Employers Under Big Apple's Anti-Discrimination Law," March 10, 2015
- "Second Circuit Finds Possible Pretext in Vague Justification for Non-Transfer," July 17, 2014
- "New Jersey Looks to Limit Use of Criminal History in Hiring Decisions," December 20, 2013
- "Proposed New York Law Would Extend State Anti-Discrimination Protections to Unpaid Interns," November 4, 2013
- "New Jersey Employers May Not "Like" State's New Social Media 'Privacy Settings,'" September 12, 2013
- "New Jersey Supreme Court Expands State Law Retaliation Claims," July 19, 2013
J.D., Georgetown University Law Center, 2008, cum laude
B.A., Hofstra University, 2005, magna cum laude
- New York
- United States Court of Appeals for the Second Circuit
- United States District Courts for the Southern and Eastern Districts of New York


text:
GOLD TEST ACID KIT
Includes the 10k, 14k, and 18k testing acids along with the 2" x 1 1/2" test stone.
HOW DO THESE QUALITY GOLD TESTING ACIDS AND ACCESSORIES WORK?
The basic gold testing procedure is to make a small scratch test of the sample metal you are testing on the test stone. Drop a few drops of the test acid on the sample and observe it. If the quality of the gold is equal to or greater than the karat quality rated on the test acid the acid will not effect the sample. If the tested metal is not gold or is of a lower purity than the quality rated on the test acid the acid will destroy the sample. An optional method is to make two scratches on the stone, one with the sample metal being tested and another with a test needle of known purity gold. When you apply the acid you observe both scratches to see how they compare.

domain: chemistry


text:
|Pardee Keynote Symposia|
|Download the Program book|
|Poster Session Map||Convention Center Floor Plans|
Geologic Consequences of Hurricane Irene — A Preliminary Assessment
Monday, 10 October, noon–1 p.m.,
Minneapolis Convention Center, Room 101A–C
Rivers and streams in the northeastern United States reached record levels as a result of Hurricane (Tropical Storm) Irene’s rainfall. More than 80 U.S. Geological Survey streamgages measured record peaks. The massive flooding that has occurred in parts of Vermont displaced geologists from the state survey office and fluvial geomorphologists in Vermont and New Hampshire have been in disaster response mode.
We have assembled experts from New Jersey, Pennsylvania, Massachusetts, New York and Vermont to provide examples of the geomorphic changes that occurred over a broad area and on a scale rarely seen. They will present data from several watersheds including hydrographs, estimated peak flows in rivers where gages were removed in order to save them from destruction, estimates of sediment moved and recurrence intervals. They will illustrate the flooding and landslides with photos and video.
This GOES-13 satellite image is of Hurricane Irene just 28 minutes before the storm made landfall in New York City. The image shows Irene's huge cloud cover blanketing New England, New York and over Toronto, Canada. Shadows in Irene's clouds indicate the bands of thunderstorms that surrounded the storm.
Image Credit: NASA/NOAA GOES Project
click on photo for larger image


text:
BACK TO NEWS
Industry News 3/1/2012
|The Action Sports Employment Market, 2012|
By Mike Lewis & Chad Mihalick (Written by Mike Lewis)
(Editor's note - click image below to download a printable PDF.)
The last few years have been a wild ride. News of layoffs have peppered our headlines, while at the same time, top-end executives have played musical chairs, bouncing from company to company chasing larger market caps, while talented middle-level execs and recent graduates languished in the unemployment lines.
The survey was conducted online and by phone, with a broad range of companies—both large and small—as well as individuals, from senior level execs to newly minted graduates, weighing in.
Analyzing the data turned up some interesting trends—perhaps the most noticeable being that most companies avow to not experiencing significant salary cuts and layoffs during the last couple of years. We sliced and diced this by company size and market cap, and it didn’t make any difference, the averages wound up just that. Whether it’s restructuring, short-term memory loss, or the fact that everyone we interviewed is still in business, most companies say they came through the Great Recession relatively unscathed.
While finding staff and keeping them has become easier for companies in a soft economy, that’s not to say that companies are taking them for granted. Employee satisfaction is still incredibly important, and as companies start ramping up hiring, the industry should continue to receive great grades from its most important customers—its employees.
EMPLOYER REPORT CARD


text:
For Authors and Editors
Types of papers
Mini reviews should be short updates (3-5 journal pages) concerning the very latest progress in any field covered by BIODEGRADATION. Please send questions related to the relevance of a particular topic to the Mini Review Editor.
Useful Resources for Authors and Editors:


text:
Want to participate in a short research study? Help shape the future of investing tools and you could win a $250 gift card!
The big shareholder groups in Animoca Brands Corporation Limited (ASX:AB1) have power over the company. Institutions will often hold stock in bigger companies, and we expect to see insiders owning a noticeable percentage of the smaller ones. I quite like to see at least a little bit of insider ownership. As Charlie Munger said 'Show me the incentive and I will show you the outcome.'
With a market capitalization of AU$140m, Animoca Brands is a small cap stock, so it might not be well known by many institutional investors. Taking a look at our data on the ownership groups (below), it's seems that institutional investors have not yet purchased much of the company. We can zoom in on the different ownership groups, to learn more about AB1.
What Does The Institutional Ownership Tell Us About Animoca Brands?
Many institutions measure their performance against an index that approximates the local market. So they usually pay more attention to companies that are included in major indices.
Institutions own less than 5% of Animoca Brands. That indicates that the company is on the radar of some funds, but it isn't particularly popular with professional investors at the moment. So if the company itself can improve over time, we may well see more institutional buyers in the future. It is not uncommon to see a big share price rise if multiple institutional investors are trying to buy into a stock at the same time. So check out the historic earnings trajectory, below, but keep in mind it's the future that counts most.
Hedge funds don't have many shares in Animoca Brands. As far I can tell there isn't analyst coverage of the company, so it is probably flying under the radar.
Insider Ownership Of Animoca Brands
While the precise definition of an insider can be subjective, almost everyone considers board members to be insiders. Company management run the business, but the CEO will answer to the board, even if he or she is a member of it.
Insider ownership is positive when it signals leadership are thinking like the true owners of the company. However, high insider ownership can also give immense power to a small group within the company. This can be negative in some circumstances.
Our most recent data indicates that insiders own a reasonable proportion of Animoca Brands Corporation Limited. It has a market capitalization of just AU$140m, and insiders have AU$24m worth of shares in their own names. I would say this shows alignment with shareholders, but it is worth noting that the company is still quite small; some insiders may have founded the business. You can click here to see if those insiders have been buying or selling.
General Public Ownership
The general public, mostly retail investors, hold a substantial 65% stake in AB1, suggesting it is a fairly popular stock. This size of ownership gives retail investors collective power. They can and probably do influence decisions on executive compensation, dividend policies and proposed business acquisitions.
Private Company Ownership
It seems that Private Companies own 14%, of the AB1 stock. Private companies may be related parties. Sometimes insiders have an interest in a public company through a holding in a private company, rather than in their own capacity as an individual. While it's hard to draw any broad stroke conclusions, it is worth noting as an area for further research.
While it is well worth considering the different groups that own a company, there are other factors that are even more important.
I like to dive deeper into how a company has performed in the past. You can access this interactive graph of past earnings, revenue and cash flow for free .
Of course this may not be the best stock to buy. Therefore, you may wish to see our free collection of interesting prospects boasting favorable financials.
NB: Figures in this article are calculated using data from the last twelve months, which refer to the 12-month period ending on the last date of the month the financial statement is dated. This may not be consistent with full year annual report figures.
We aim to bring you long-term focused research analysis driven by fundamental data. Note that our analysis may not factor in the latest price-sensitive company announcements or qualitative material.
If you spot an error that warrants correction, please contact the editor at firstname.lastname@example.org. This article by Simply Wall St is general in nature. It does not constitute a recommendation to buy or sell any stock, and does not take account of your objectives, or your financial situation. Simply Wall St has no position in the stocks mentioned. Thank you for reading.

domain: finance


text:
Oosthuizen 12/1 to bounce back!
Louis Oosthuizen is straight back out after his second place at the Masters and Hills rate him as their 12/1 joint favourite (with Martin Kaymer) to win in Malaysia.
Luke Donald is back in action at the Heritage and he can be backed at 6/1 with Matt Kuchar a 16/1 shot.
Rory Mcllroy is taking a break for a few weeks and despite his disappointing form at the Masters, Hills rate him as their 7/1 joint favourite to win the second major of the year (US Open).
"There are a lot of question marks after what was a strange Masters, Louis is going to be emotionally drained & Donald could be doubting his swing," said Hill's spokesman Rupert Adams.
Maybank Malaysian Open: 12/1 Martin Kaymer, 12/1 Louis Oosthuizen, 12/1 Charl Schwartzel, 16/1 Nicolas Colsaerts, 16/1 Matteo Manassero, 22/1 George Coetzee, 25/1 Soren Kjeldsen, 33/1 Alvaro Quiros 33/1 Mohd Sidd
RBC Heritage: 6/1 Donald, 16/1 Kuchar, 18/1 Furyk, 20/1 Bo Van Pelt, 22/1 Webb Simpson, 25/1 Ernie Els, 28/1 Brandt Snedeker, 30/1 Aaron Baddeley, Bud Cauley, 30/1 Kevin Na, 33/1 Padraig Harrington, 35/1 Jeff Overton 40/1 Jason Dufner
112th US Open: 7/1 Rory Mcllroy, 7/1 Tiger Woods, 10/1 Phil Mickelson, 16/1 Lee Westwood, 16/1 Luke Donald, 33/1 Bubba Watson, 33/1 Hunter Mahan, 33/1 Martin Kaymer, 33/1 Adams Scott, 40/1 Nick Watney, 40/1 Padraig Harrington, 40/1 Steve Stricker, 40/1 Jason Day, 40/1 Sergio Garcia, 40/1 Justin Rose, 50/1 Bar

domain: sports


text:
Frenchman now expected to search for a new club.
AC Milan midfielder Mathieu Flamini has failed to reach an agreement on a contract extension after his previous deal expired earlier this week.
The 29-year-old enjoyed an improved season last year, adding goals to the Rossoneri’s cause as they fought to the final day of the season to secure Champions League football.
However, it wasn’t enough for the Frenchman to be offered a significantly improved new deal as his old contract expired on Monday, with a difference in wages and duration causing a conflict.
It is understood that Flamini had requested a higher salary whilst also pushing for an additional year to the contract offer, demands Milan were not willing to cater for.
Vice-president Adriano Galliani is said to have been willing to offer a new two-year deal, but that was not enough for Flamini who eyed a lengthier stay given his age.
As a result, the former Arsenal man will now begin the search for a new club, and isn’t likely to make too many friends at Milan by revealing to a reporter that he wouldn’t be against signing for city rivals Inter.
However, following on from his representative making it clear that he is unlikely to agree to Milan’s deal, Flamini has himself confirmed he is without a club for the time being.
“I’m not going to accept the offer of only two years,” he told Corriere dello Sport as reported by football-italia.net.
“At 29, after five season with the Rossoneri, and after the four important goals I scored last season, I deserved diffferent treatment.
“I am disappointed, and I’m going my own way.
“My future? Anything is possible.”
Flamini was clearly disappointed with the treatment he has been handed, but nevertheless it would appear the Rossoneri will now step up their interest in Sampdoria’s Andrea Poli, with the 23-year-old expected to sign this week.


The Right to Rent scheme – by which landlords or their agents must check the immigration status of tenants and evict any tenant who does not have right to live in the UK – is likely to go live nationally by next April, and possibly much sooner.
There could be a phased roll-out across England from this autumn onwards.
Landlords – and presumably their agents – who do not comply face fines or prison sentences of up to five years.
The eviction of illegal tenants will be abrupt, and without having to go through court.
It would follow the issuing of a notice by the Home Office when an asylum application fails, confirming that the tenant no longer has the right to rent.
The Government is expected to enact new criminal offences as early as next month. Normally, measures enacted in September come into force the following April. However, in view of the crisis in Calais, sources say there is speculation that ministers could decide to bring implementation sharply forward.
Greg Clark, the communities secretary, said the legislation will also create a blacklist of persistent rogue landlords and letting agents to allow councils to know where to concentrate their enforcement action.
“We are determined to crack down on rogue landlords,” said Clark.
There will also be measures to prevent the letting out of sub-standard properties.
The new measure looks to be controversial.
The pilot scheme in the West Midlands has been running only since December and awaits evaluation.
In the pilot, there is no criminal penalty, with civil sanctions of up to £3,000.
Also in the pilot, landlords are able to assign Right to Rent responsibilities to their agents, and it is thought that this same system will continue in the national scheme.
The new clampdown is already raising fears that landlords and agents will simply discriminate against certain types of prospective tenants – including those with a right to live in the UK.
The Joint Council for the Welfare of Immigrants said that the pilot has shown serious shortcomings, with British people who have foreign accents finding it difficult to find somewhere to rent.
Lawyer and policy director of the Residential Landlords Association David Smith told the BBC’s World at One that there was evidence that landlords in the pilot were reluctant to let their properties to anyone without a valid passport.
He said: “This means that huge segments of the population, including genuine UK national who do not have passports – of whom there are many – are being excluded.”
There are also accusations that the Government is guilty of a knee-jerk reaction to the Calais crisis.
However, David Cox, managing director of ARLA, said: “ARLA believes that the measures announced by the Government today are a good first step and we welcome the proposals in principle.
“The plans will help to weed out the minority of rogue landlords who exploit vulnerable immigrants for their own financial gain and, with the introduction of a new five year imprisonment penalty, will help to deter other such unscrupulous individuals from entering the private rented sector.
“The proposals also build upon the Right to Rent checks as imposed by the Immigration Act 2014.
“We will be organising training sessions for our members to ensure they are fully prepared and understand the new rules and we urge all letting agents to ensure they are ready for the impending roll out.”


text:
Comfort Inn & Suites Tuscumbia - Muscle Shoals - Muscle Shoals, Florence
5101 Hwy 43 S - Florence1-877-662-6988
Welcome to Comfort Inn & Suites Tuscumbia - Muscle Shoals!
This Muscle Shoals hotel offers free Wi-Fi, gift shops/newsstands and an elevator (lift). You will want to pack your swimsuit and towel as this hotel features a seasonal outdoor pool. If you are visiting United States of America on vacation or for business, make sure to take some time to relax as you will find a fitness center on site, so make the most of it!
Sit back and relax in the lobby which has free newspapers and coffee. Other amenities include laundry facilities.
We show TripAdvisor reviews for new Venere hotels that haven't been reviewed yet
Check Availability at Comfort Inn & Suites Tuscumbia - Muscle Shoals
Breakfast at Comfort Inn & Suites Tuscumbia - Muscle Shoals
A free buffet breakfast is served each morning.
- Laundry room
- Gift shop
- Outdoor swimming pool
- Front desk - 24 hour
- Internet/Email services
- Free parking
Check-in: after 3 PM.
Check-out: before 11 AM.
Government-issued photo identification and a credit card or cash deposit are required at check-in for incidental charges.
Special requests are subject to availability upon check-in and may incur additional charges. Special requests cannot be guaranteed.
Comfort Inn & Suites Tuscumbia - Muscle Shoals has 62 guest rooms, all featuring free local calls and free Wi-Fi. Kitchen amenities include refrigerators and microwaves, making it possible to dine in while you travel.
Business travelers will have access to desks and phones during their Muscle Shoals visit. Additional guest comforts include air conditioning and coffee/tea makers.
Bathrooms are outfitted with hair dryers.
- Air conditioning
- Mini fridge
- Coffee and tea maker
- Hairdryer in each room
- In-room ironing facilities
- Wireless Internet access
Distances from Comfort Inn & Suites Tuscumbia - Muscle ShoalsIvy Green - 4.4 km / 2.7 mi
Indian Mound and Museum - 8.5 km / 5.3 mi
Frank Lloyd Wright Rosenbaum House - 8.9 km / 5.5 mi
W.C. Handy Home - 9.3 km / 5.8 mi
Shoals Community Theater - 9.8 km / 6.1 mi
Kennedy-Douglass Center for the Arts - 10.1 km / 6.2 mi
Pope's Tavern Museum - 10.2 km / 6.3 mi
University of North Alabama - 10.5 km / 6.6 mi
Shoals Conference Center - 10.8 km / 6.7 mi
Renaissance Tower - 10.9 km / 6.8 mi
Regency Square Mall - 14 km / 8.7 mi
Children's Museum of the Shoals - 14.3 km / 8.9 mi
Joe Wheeler State Park - 34.4 km / 21.4 mi
The preferred airport for Comfort Inn & Suites Tuscumbia - Muscle Shoals is Muscle Shoals, AL (MSL-Northwest Alabama Regional) - 5.7 km / 3.5 mi.
Distances are calculated in a straight line from the property's location to the point of interest or airport and may not reflect actual travel distance.
Distances are displayed to the nearest 0.1 km and mile.
Transportation services Comfort Inn & Suites Tuscumbia - Muscle Shoals
- Free parking

domain: travel


text:
Robert H. Miller, PhD, a neuroscientist, researcher, and administrator in the GW School of Medicine and Health Sciences, has been named the university’s new vice president for research.
The GW School of Medicine and Health Sciences renamed the Department of Anatomy and Regenerative Biology to the Department of Anatomy and Cell Biology.
Adam Friedman, MD, professor of dermatology at the GW School of Medicine and Health Sciences, and his team investigated the use of nanotechnology to improve efinaconazole treatment and make it more cost effective.
A recent clinical trial at GW showed no significant support for tamsulosin use for kidney stone passage.
Research published in Cancer Cell by George Washington University (GW) Cancer Center researcher Alexandros Tzatsos, MD, PhD, found an important connection between the KDM6A gene and the most aggressive form of pancreatic cancer.
A team of researchers at the GW School of Medicine and Health Sciences compared the toxicological impact of cryosurgery with an FDA-approved topical 40 percent hydrogen peroxide solution (A-101) for the treatment of seborrheic keratosis, in human...
Researchers at GW and the University of Georgia are partners in a project that will soon be able to provide a way for questions asked by those studying glycoscience to be answered by big data.
A research team led by Brad Jones, PhD, from GW School of Medicine and Health Sciences found that HIV-infected cells are resistant to CD8+ T-cells. The research was published in the Journal of Clinical Investigation.
Researchers at Baylor College of Medicine, in collaboration with a team of researchers at the George Washington University and the René Rachou Institute, have received funding from the National Institutes of Health for a Phase Ib clinical trial...
Kenneth Harwood, PhD, PT, associate professor of clinical research and leadership, received the Health Services Research Pipeline Grant from the Foundation for Physical Therapy to study how the timing of physical therapy (PT) care and provider...