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In what will seem like no time at all, your child will morph from a toddler and into a teenager. While many physical, emotional, and mental changes will occur, so will their habits, more specifically, their brushing, flossing, and rinsing routine. It might appear as if they don't need to change how they care for their teeth but as soon as those baby teeth fall out and are replaced with permanent, adult teeth, the game changes. Hear from a dentist in Alamo Ranch who can explain what changes will likely occur with each developmental stage your child enters and what you can do to help them along the way.(more…)
Alamo Springs Dental Blog
From Toddler to Teen: How Your Child's Dental Habits Change with Age
July 15, 2020
A Dentist Answers 5 Common Concerns of Patients Most At-Risk for COVID-19
July 7, 2020
Are you preparing to see your dentist in Alamo Ranch? Do you worry because you are considered "at-risk" that you might contract a harmful virus that could cause you to become ill? Will you be okay if you postpone your dental checkup and cleaning another few months? You are not alone in wondering about your health when leaving the house. This is why your dentist is going above and beyond to not only provide a safe and secure environment for all patients, but they are also taking the time to address the common concerns of individuals most at-risk for contracting COVID-19. Read on to find out what your dental professional has to say.(more…)
5 Ways Your Dentist is Keeping Your Health and Safety in Mind
June 13, 2020
Are you ready to see your dentist in Alamo Ranch for a regular checkup but are still worried about the cleanliness of their office? You're not alone in your thinking; however, your dental team doesn't want this concern to keep you from getting the care you need. But how can you know it's safe? To offer you reassurance and better peace of mind, here are 5 ways your dentist is preparing your upcoming visit.(more…)
How to Support Equality While Living in San Antonio
June 8, 2020
As if the last few months have not been difficult enough with COVID-19 spreading rapidly throughout the world, the recent events surrounding racial injustice are causing people all over the globe to rise and stand in solidarity with the black community. With protests occurring in cities throughout the United States, many individuals are wondering how else they can show they care. A recent article in San Antonio Magazine shared a list of black-owned businesses to support in San Antonio, one of which is your very own Alamo Springs Dental! If you're interested in spending your money at other minority-owned establishments, here are a few to consider.(more…)
Why Should I Rinse My Mouth Before My Dental Appointment?
May 3, 2020
Brushing and flossing are great ways to minimize harmful bacteria within your mouth, but did you know that rinsing with an antimicrobial mouthwash also has many great benefits? Whether you're accustomed to using one of the many products on the market to flush out left-behind particles or you are new to the idea of rinsing out your mouth, your dentist in Alamo Ranch strongly recommends this step before arriving for your dental appointment. Spend some time reading the following article to learn how mouthwash can keep you and others safe amid the COVID-19 (coronavirus) pandemic.(more…)
4 Effective Ways to Maintain Good Oral Hygiene During COVID-19
April 3, 2020
Whether you are practicing social distancing or undergoing self-quarantine, COVID-19 is certainly making life interesting for people throughout the world. Since you are likely staying at home more, and since most dental offices are closed and only seeing emergency patients, you may be thinking about your oral hygiene routine and ways to improve it. If you're wondering how you can be keeping up with your dental hygiene during quarantine, check out these four helpful tips that will give you increased confidence in your ability to maintain a healthy smile.(more…)
Understanding Bruxism: 4 Dangers of Teeth Grinding
March 5, 2020
Do you grind your teeth at night? When stressed, do you find yourself clenching and putting pressure on your jaw? These common habits are easy to form, yet hard to break. Also known as bruxism, millions of adults and children experience its negative effects, some of which are irreversible. Whether you or someone you love suffers from this particular dental problem, spend some time learning more about the dangers of teeth grinding and what you can do to achieve relief and save your smile.(more…)
Why You Should Start the Year Off Right with Dental Checkups and Screenings
January 3, 2020
If you're the kind of person who likes to start fresh each year, complete with a list of resolutions, why not start the year off right with dental checkups and screenings? Because the mouth and body are closely interconnected, keeping your teeth, gums, and overall oral health in optimal condition is vital to prevent additional health problems in the future. If you're ready to take charge and commit to improving your smile and well-being, read on to learn why regular checkups and screenings are so important.(more…)
Dental Crown vs. Dental Bridge: What You Need to Know
December 3, 2019
Do you know the difference between a dental crown vs. a dental bridge? For years, dentists have continually turned to these reliable and effective restorations as a way to restore smiles and improve oral health. Whether it is a missing tooth or one that is severely decayed, these custom-made prosthetics can give you a new lease on life as well as boost your confidence like never before. Read on to learn more about these useful restorations and find out which one is right for you.(more…)
Why Are 6 Month Checkups Important in Alamo Ranch? Let a Dentist Explain
September 2, 2019
Have you asked yourself, "Why are 6-month checkups important in Alamo Ranch?" It's doubtful you're alone in asking that question, but let us assure you, there are many reasons you should adhere to this common timeline. From cavities to gum disease, there are many reasons your dentist wants to see you twice a year for a checkup and cleaning. Find out what you can expect during these appointments and what problems can be avoided by keeping your dental visits a priority.(more…)


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I have a single pole circuit and was wondering if the add-on switches must be wired to the master switch or just have power? I have a multi-gang box in the bathroom and could install the add-on switch in this box but it is not wired to the bedroom light (a single pole circuit.)
If you're using the common GE/Jasco/Honeywell, the add-on switches don't work independently. They communicate with the master switch over the wired connection.
If you want control from a second location, you could put another smart (master) switch in this location without a load and connect it programmatically, or use another button / scene controller.
(edited for clarity per JD's comments below)
It depends on the exact brand/model of the switches that you are using. The GE, Inovelli, and Leviton three-way set up do use physical traveler wires. But many of the other brands including Linear/GoControl, Eaton Cooper, Homeseer, and Lutron Use "virtual" three-way setups where both the master and the auxiliary switch have a radio. The auxiliary sends a message to the hub and the hub sends a message to the master. No physical wires are required between them, they can even be on completely opposite sides of the house. There are pros and cons to both approaches, so it just depends on what you need for each specific switch location.
Example of a virtual three-way add on switch.


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Your father moved to another country, so you decide to move back to the city where you were born. There, you will discover friendship, love and lots of sex. Dawn of Malice has a unique feature, you can choose your moral alignment at the beginning, and this will allow you to have some UNIQUE options during your gaming. Think carefully, because there is no going back from your actions in this game.
Adult Game Updated: 29-05-2020
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Endotoxin tolerance is characterized by the suppression of further TNF release upon recurrent exposure to LPS. This phenomenon is proposed to act as a homeostatic mechanism preventing uncontrolled cytokine release such as that observed in bacterial sepsis. The regulatory mechanisms and interindividual variation of endotoxin tolerance induction in man remain poorly characterized. In this paper, we describe a genetic association study of variation in endotoxin tolerance among healthy individuals. We identify a common promoter haplotype in TNFRSF1B (encoding TNFR2) to be strongly associated with reduced tolerance to LPS (p = 5.82 × 10−6). This identified haplotype is associated with increased expression of TNFR2 (p = 4.9 × 10−5), and we find basal expression of TNFR2, irrespective of genotype and unlike TNFR1, is associated with secondary TNF release (p < 0.0001). Functional studies demonstrate a positive-feedback loop via TNFR2 of LPS-induced TNF release, confirming this previously unrecognized role for TNFR2 in the modulation of LPS response.
The inflammatory response to LPS, an invariant component of the outer membrane of Gram-negative bacteria, is mediated via a TLR-dependent transduction cascade (1, 2). This process is tightly regulated and characterized by the coordinate expression and release of multiple cytokines including TNF, IL-6, and IL-1β. Much of the early morbidity associated with Gram-negative bacterial infections is attributable to LPS-induced TNF release (3). Therefore, mechanisms controlling this response in man have vital implications in health and disease. Repeated or chronic exposure to LPS, or indeed other TLR ligands, leads to rapid tolerance to their proinflammatory actions both at a cellular level and at the level of the organism. This tolerance to LPS is referred to as endotoxin tolerance and was described over a century ago (4, 5). However, only recently have the underlying molecular mechanisms begun to be elucidated (6). Endotoxin tolerance is now recognized as a multifactorial process characterized by the induction of negative regulators of TLR signaling that can be proteins such as SHIP (7) and IL-1R–associated kinase-M (8) or microRNAs including micro (mi)R146, miR155, and miR9 (9, 10). Concomitant to this, chronic exposure to LPS leads to chromatin remodeling and the selective silencing of proinflammatory genes, resulting in a more profound state of tolerance (11).
The multiple conserved biological mechanisms of tolerance across species underline the importance of homeostatic regulation of innate immunity. In animal models, induction of endotoxin tolerance can render lethal doses of LPS sublethal, primarily by restricting further LPS-induced TNF release. Failure to silence innate immune activity is implicated in many acute and chronic inflammatory states, ranging from sepsis and autoimmune conditions to cancer. Identification of mechanisms regulating innate immune tolerance in man provides targets for therapeutic intervention to silence dysregulated innate immune responses.
Genetic polymorphisms play a significant role in determining interindividual variation in cytokine responsiveness to LPS (12, 13), but the phenotype of endotoxin tolerance has not been characterized in this respect. Genome-wide association analyses have demonstrated the impact of common genetic variants on multifactorial traits, highlighting the importance of noncoding sequence variants that modulate gene expression (14). For a number of common diseases such as type 1 diabetes and Crohn's disease, genome-wide association studies have isolated genes for which encoded products had not been previously considered in disease pathophysiology (15). In this study, we sought to define the extent of variation in endotoxin tolerance to repeated stimulation with LPS based on TNF release among healthy volunteers. We aimed to identify functional genetic variants associated with tolerance induction and, in doing so, gain novel insights into the regulation of tolerance induction in man. We show that a common haplotype spanning TNFRSF1B, the gene encoding TNFR2 (denoted by its alternative gene name TNFR2 in the following text), is strongly associated with reduced tolerance to LPS. We proceed to demonstrate that this haplotype is associated with differential nuclear factor binding and is specifically associated with increased basal expression of TNFR2. We subsequently show that LPS-induced TNF activity can positively feedback through TNFR2, impacting upon the duration and magnitude of TNF release.
Materials and Methods
Ethics statement
This study was approved by the Oxfordshire Research Ethics Committee (REC reference 06/Q1605/55). All volunteers gave written informed consent.
Study volunteers, PBMC purification, and LPS tolerance assays
The recruitment and demographics of the healthy volunteer cohort, sample collection, PBMC purification, and cell culture conditions have been previously described (13). In summary, healthy volunteers without a history of recent viral or bacterial infection were venesected in the morning and PBMCs purified using a Ficoll-paque PLUS (GE Healthcare, Chalfont St Giles, U.K.) gradient within 2 h. Cells were counted with a hemocytometer and resuspended at 2.5 × 106 cells/ml in RPMI 1640 growth medium (Sigma-Aldrich) supplemented with l-glutamine (2 mM), penicillin/streptomycin (100 U/ml penicillin, 0.1 mg/ml streptomycin), and 10% v/v heat-inactivated gold-standard FCS, with three biological replicate assays per condition set up for each volunteer. After overnight incubation, cells were washed in fresh media then resuspended in media alone or with LPS at 2 ng/ml (L-4391, lot 114K4133; Sigma-Aldrich), for 6 h prior to separation of cells from supernatant by centrifugation. To assess tolerance, additional naive or LPS-treated cells (2 ng/ml for 6 h) were exposed to LPS (20 ng/ml) for a further 6 h. Three biological replicate assays per condition were set up for each volunteer and the mean used in subsequent statistical analysis. Supernatants for ELISA were stored at −80°C prior to quantification; cell pellets for RNA extraction were resuspended in RLT buffer (Qiagen, Valencia, CA) supplemented with 2-beta-mercaptoethanol. Where indicated, a TNFR2 allosteric modulator Ab (anti-human TNF-R II clone 80M2, catalog number HM2022; Hycult Biotech, Uden, The Netherlands) or mouse IgG1 isotype control (functional-grade) mAb (catalog number M075-3M2; Medical and Biological Laboratories, Nagoya, Japan) was added to cultures at a concentration of 2.5 μg/ml for the times indicated. For the experiments involving purified monocytes, CD14+ monocytes were positively selected with Miltenyi CD14+-labeled microbeads (catalog number 130-050; Miltenyi Biotec, Bergisch Gladbach, Germany) according to instructions, with all steps performed at 4°C or on ice. Experiments with monocytes were carried out at the lower concentration of 5 × 105 cells/ml.
ELISAs were performed on all samples in duplicate as described (13
RNA extraction, cDNA synthesis, and quantitative PCR
Total RNA was prepared using the RNeasy Mini kit (Qiagen) as described with cDNA prepared with SuperScript III (Invitrogen, Paisley, U.K.) using random hexamers (13). Quantitative PCR was carried out using SYBR Green Supermix (Bio-Rad, Hemel Hempstead U.K.) on the CFX96 C1000 Thermal Cycler (Bio-Rad). PCR efficiency was determined and melt curve analysis performed for gene-specific primer sets (Supplemental Table I). Relative gene transcript levels were determined by the ΔΔCt method.
Genotyping, haplotype, and statistical analysis
The TNFR2 variable number tandem repeat (VNTR) was genotyped by amplification of a 150-bp fragment by PCR (primer sequences shown in Supplemental Table I), and the number of repeats was directly identified from electrophoresis of a 3% agarose gel. Genotyping of the volunteer cohort using the human CVD bead array chip (Illumina, San Diego, CA), computation of linkage disequilibrium (LD) and haplotypes using the HaploView 3.3.2 program (16), and expression quantitative trait mapping using PLINK (17) was performed as previously described (13) with verification using SPSS (SPSS), R package, and SNPTest. Standard quality control measures were used with exclusion criteria of maximum per single nucleotide polymorphism (SNP) missing (geno >0.1 and maf <0.03). The genotyping rate was >98.8%. For each SNP analyzed, the phenotypic mean for the three genotypic states was compared using the Wald test statistic to generate a p value that does not require that the data fit a normal distribution. Covariates age, ethnicity, and sex were included in PLINK analysis to further interrogate observed associations. Nonparametric statistics and log transformation were applied where data were not normally distributed, and otherwise unpaired t tests were used for analysis of expression data for specific SNPs, data passing tests of normality. Permutation analysis was performed using a label-swapping procedure and an adaptive algorithm. Analysis was performed with one million permutations using a within-cluster algorithm as described (13). The empirical p value is robust with respect to normality of phenotype and multiple testing issues. All correlations were carried out using linear regression least-square fit models with statistical relevance and correlation coefficients described for each association. Where indicated, two-tailed t tests were used.
Nuclear extracts and EMSAs
Nuclear extracts were prepared, oligonucleotide probes were radiolabeled with [32P]deoxycytidine 5′-triphosphate (PerkinElmer, Beaconsfield, U.K.) by fill-in of 5′ overhanging ends using Klenow and unlabeled 2'-deoxyadenosine 5′-triphosphate, deoxyguanosine 5′-triphosphate, and deoxythymidine 5′-triphosphate, and EMSA performed as previously described (18, 19). Probes were generated by annealing forward and reverse oligonucleotides. Sequences are shown in the Supplemental Table I.
Variation in endotoxin tolerance at the level of TNF release in response to LPS
Similar to previous studies (20), we noted that a 6 h pretreatment of cells with LPS established marked tolerance to further LPS-induced TNF release. We also observed a degree of interindividual variation at this time point, permitting investigation of underlying genetic contributions to this. We therefore based our assay on TNF release in response to LPS stimulation for 6 h comparing naive versus pretreated cells. We were interested in the individual silencing of TNF production after LPS treatment, and so to normalize for variation in initial response, due to either interindividual variation in PBMC composition or other intrinsic sensitivities to LPS, values were expressed relative to the naive response. This approach allowed us to focus upon silencing of TNF responses in a sample most representative to that in circulation. We proceeded to recruit a cohort of 96 healthy volunteers and to define variation in tolerance to LPS based on three replicate stimulation assays for a given volunteer on a particular day, with the mean value used in subsequent analyses.
Over a 6-h period, LPS treatment (2 ng/ml) resulted in a pronounced release of TNF into cell culture supernatants, with significant interindividual variability in induced TNF release (geometric mean 664 pg/ml; 95% confidence interval [CI] 593–743 pg/ml; range 118–3511 pg/ml) (Fig. 1). Cells were subsequently washed extensively and restimulated with a 10-fold higher LPS concentration (20 ng/ml) for a further 6 h. In tandem with this, naive cells that had been treated identically (washed in same manner), but had been exposed to media alone, were stimulated with LPS at the higher concentration. Cells pre-exposed to LPS showed a pronounced reduction in secondary release of TNF upon restimulation and also showed significant variability across the cohort (geometric mean 83 pg/ml; 95% CI 69–100 pg/ml; range 11–550 pg/ml) (Fig. 1).
Although most individuals showed a high level of silencing on secondary stimulation (mean 15.8%; SD 10.8; range 1.4–59), several individuals showed reduced induction of tolerance with secondary responses 30–50% that of their primary response (Fig. 1). Interestingly, we did not note an association between the magnitude of the primary and secondary TNF response (data not shown). When we analyzed the effect of secondary stimulation on IL-6 production at 6 h, we did not see tolerance, with most individuals releasing a similar amount or more IL-6 (Fig. 1), confirming continued cell viability and reflecting mechanistically different silencing mechanisms between TNF and other acute-phase cytokines at this relatively early time point in endotoxin tolerance.
Common genetic markers associated with reduced endotoxin tolerance
To determine genetic modulators of endotoxin tolerance, we proceeded to map tolerance as a quantitative trait among the cohort of 96 healthy volunteers. Individuals were genotyped at 45,237 SNPs using the Illumina HumanCVDv1 beadchip (Illumina), which has high density coverage of ∼2000 genes implicated in immune and inflammatory responses of specific relevance to vascular pathology, metabolic disorders, and inflammatory disease states (21).
We found the most significant associations were with a cluster of SNPs at or near to TNFR2 at chromosome 1p36.22 (Fig. 2). Moreover, analysis of quantile–quantile plots highlighted that SNPs in TNFR-related genes or TNFR-associated factor (TRAF) family genes accounted for the majority of the overdispersion of p values over the expected in terms of statistical association (Fig. 2). Three SNPs in complete LD upstream of the TNFR2 gene (rs522807, rs5745938, and rs625847) showed the strongest association with endotoxin tolerance; the mean tolerance was 31.3% (95% CI 17.2–45.5) among individuals possessing a copy of the haplotype defined by rs522807 compared with 14.4% (95% CI 12.5–16.3) among those without (Supplemental Fig. 1). By contrast, we found no association with initial TNF release (Fig. 2). The reduced tolerance-associated SNP rs522807 shows marked differences in allele frequency among different populations worldwide. The tolerance-associated A allele (denoted as the ancestral allele as this is the allele observed in chimp, orangutan, and macaque genomes) was present at a frequency of 8% in northern European-derived populations, absent in Asian populations but present at a frequency of near 50% in equatorial Africa (Supplemental Fig. 2).
rs522807 is associated with increased expression of TNFR2 in the basal state
Given the association of SNPs spanning TNFR2 with endotoxin tolerance, we sought to define if the same SNPs were modulating expression of this gene locally in a cis-acting manner. Using data from real-time quantitative RT-PCR, we proceeded to carry out an expression quantitative trait analysis for TNFR2. Interestingly, we found that rs522807 marked an eQTL for TNFR2 (p = 4.9 × 10−5). Individuals possessing a copy of the endotoxin-associated SNP rs522807 had significantly increased basal expression of TNFR2 (Fig. 2, Supplemental Fig. 1). This indicated that the promoter polymorphisms associated with decreased tolerance enhanced basal expression of the receptor, suggesting the hypothesis that irrespective of genotype, basal expression of TNFR2 positively influences secondary responses by regulating tolerance.
TNFR2 expression correlates with secondary, but not primary, TNF release
We proceeded to test this hypothesis by investigating whether there was any correlation between the expression of TNFR2 and LPS-induced TNF response. We postulated that feedback of LPS-induced TNF through basally expressed TNFR2 may influence the secondary response, although we were uncertain whether this would contribute to desensitization and tolerance or have the opposite effect. Using real-time quantitative RT-PCR, we found there was no association between basal expression of either TNFR1 or TNFR2 and primary release of TNF (data not shown). However, we found a highly significant correlation between the basal expression of TNFR2 mRNA and secondary TNF protein release (p < 0.0001) (Fig. 3). LPS stimulation resulted in a robust induction of TNFR2 expression (4.8-fold induction; 95% CI 4.2–5.4), but whereas an association between induced TNFR2 and tolerance remained, this was far more modest (r2 = 0.09; p = 0.002). This implied that the level of basal expression is of greater biological relevance in the determination of later responses. Notably, the association with tolerance was not seen for TNFR1 (Fig 3), suggesting this is a TNFR2-specific phenomenon. Analysis of expression levels of TNF mRNA rather than protein gave similar results (Supplemental Fig. 3).
Fine mapping association at TNFRSF1B and analysis of transcription factor binding
To fine map the causative functional genetic variant, we resequenced a 3.5-kb region of the TNFR2 promoter that included rs522807 (chr1: 12,146,659–12,149,575). In addition to the three single nucleotide substitutions identified from the array (rs522807, rs5745938, and rs625847), a further single nucleotide substitution (rs520916) and insertion (rs35752907) were found to be in complete LD with the haplotype defined by rs522807. Additionally, we noted a previously described VNTR comprising a 15-bp insertion within the promoter that exists in two allelic forms (either one or two repeats) (22) that has been associated with both infective and autoimmune disease processes (23, 24), although as yet it is unclear whether it influences expression of TNFR2. To answer this question, we genotyped this VNTR and found a weak association between possession of a single copy of the VNTR and TNFR2 expression, but this effect was lost when the analysis was conditioned on rs522807.
To determine if any of the SNPs within the endotoxin tolerance-associated haplotype modulated allele-specific protein–DNA interactions, EMSAs were performed. Radiolabeled oligonucleotides corresponding to each SNP allele were incubated with nuclear extracts from the human leukemic monocyte lymphoma cell line U937. We observed that rs522807 showed markedly different binding profiles between alleles using nuclear extracts from U937 cells that were specific on competition EMSA (Fig. 4). In silico analysis shows that the associated A allele of rs522807 introduces a cAMP response element (CRE) binding site, and competition experiments demonstrated that the upper allele-specific complex I is specifically competed by an oligonucleotide containing a CRE binding site, but the identity of the allele-specific CRE binding protein recruited at rs522807 remains to be defined.
As this study neared completion, data from the 1000 Genomes Project (http://www.1000genomes.org) became available. This shows that the haplotype tagged by rs522807 extends for 15.9 kb upstream of TNFR2 with 17 SNPs in complete LD (Supplemental Table II), which will require further characterization to define the causative regulatory variant(s).
Allosteric modulation demonstrates a role for TNFR2 in augmenting LPS responsiveness
Our data suggest that the expression of TNFR2 plays a direct role in modulating tolerance to LPS-induced TNF release. However, the overriding effect of increased basal expression of TNFR2 was unclear. TNFR2 is cleaved at the cell surface by ADAM 17 (ADAM metallopeptidase domain 17), a disintegrin and metalloproteinase also referred to as TACE (TNF-α converting enzyme) to form soluble TNFR2 that can antagonize the activity of TNF at its receptors with important functional consequences (25, 26). It was therefore possible that the predominant effect of overexpression of TNFR2 would be to antagonize the action of TNF. To dissect out the potential effect of TNFR2 activity in the development of endotoxin tolerance, we took advantage of an allosteric modulating mAb raised against this receptor (anti-human TNF-R II clone 80M2). Although 80M2 has no independent agonist activity, it markedly increases the affinity for TNF to TNFR2 and thus confers enhanced TNFR2 signaling in the presence of TNF only (27).
Prior to LPS application, cells were treated with 80M2 or isotype control IgG1 for 1 h before extensive washing to remove any unbound Ab. Cells were then treated with LPS for up to 10 h, with supernatants and RNA harvested in the absence of LPS and at 2, 6, and 10 h. Pretreatment with Abs did not elicit TNF release in the absence of LPS. When LPS was applied, 80M2 had no effect on early induced TNF release at 2 h, but at later time points, cells pretreated with 80M2 showed significantly enhanced LPS-induced TNF release (Fig. 5). Furthermore, expression of TNF, which was maximal at 2 h, was not significantly different from IgG1. However, at 6 h, there was delayed normalization in TNF expression (Fig. 5).
In addition to modulating the primary response to LPS, we were keen to investigate whether 80M2 could modify secondary responses to LPS and hence tolerance. Use of 80M2 in crude PBMC populations had variable effects, with cells from some individuals showing pronounced alteration in tolerance, whereas others were not affected (data not shown). We reasoned that this may be secondary to varying cellular composition of PBMC fractions. 80M2 has been shown to have activity at specific T cell populations (28), and it was reasoned that activity of the Ab at nonmonocyte populations may confound effects on tolerance. To avoid this possibility, we purified CD14+ monocytes from the PBMC fraction of unrelated healthy volunteers, thus focusing on the cells mainly responsible for LPS-induced cytokine release. Similar to the PBMC fraction, we found pretreatment with 80M2 had no independent agonist activity (data not shown), but greatly enhanced LPS-induced TNF release (2363 ng versus 1010 ng; n = 4, p < 0.05) (Fig. 5). After 6 h of 2 ng/ml LPS, monocytes were washed and restimulated for a further 3 h with 20 ng/ml LPS before measuring secondary responses. We found that this pretreatment significantly enhanced secondary TNF release in response to LPS, demonstrating activity through TNFR2 can modulate tolerance (Fig. 5). These results supported the hypothesis generated from the genetic screen whereby LPS-induced TNF acts to further amplify late release of TNF in a TNFR2-specific manner. These data show that manipulation of basally expressed TNFR2 can dictate the both the magnitude and durability of LPS-induced TNF release.
Although the phenomenon of endotoxin tolerance has been described for over a century, the biological mechanisms underlying this critical immunological process are only now beginning to be elucidated. These include the concomitant upregulation of negative regulators of TLR signaling, induction of miRNAs, which specifically target products of the NF-κB pathway, and the selective silencing of multiple proinflammatory genes. Most data emanate from mouse models, and there is a degree of discrepancy between genes implicated in the induction of endotoxin tolerance in animal studies and in man such that only IL-1R–associated kinase-M has consistently been demonstrated to play a role in both mice and men (8, 20). The significance of a more comprehensive understanding of the basis of endotoxin tolerance is underlined by the clear role for this phenomenon in the pathogenesis of diseases such as sepsis. The tolerant state is associated with increased susceptibility to subsequent infections and indeed may reflect an underlying proinflammatory but paradoxically immunosuppressed state (20). Nonetheless, the highly conserved nature of tolerance and the multiplicity of mechanisms whereby it can be achieved indicate it plays a key role in the regulation of the innate immune response. Indeed, it is postulated that endotoxin tolerance has evolved to act as a protective mechanism to prevent the cytokine storm associated with onset of sepsis and strongly associated with shock (6).
In this study, we were interested in addressing the degree to which individuals tolerized to LPS over a short period, thus reflecting silencing of TNF release. We specifically chose a 6-h pretreatment duration because in a clinical setting, the onset of sepsis can often be abrupt, with patients deteriorating rapidly over such a time period. Although studies of tolerance over 24 h show universal silencing of most inflammatory cytokines (11), we find there is a large degree of interindividual variability in the silencing of TNF over 6 h, and therefore, this time point is more applicable to an association study. It is of note that in our study we found little silencing of the IL-6 response, illustrating both the differential regulation of these genes and also the continued viability of cells.
Our interest in identifying genetic markers of endotoxin tolerance has been driven by the observations that responses to innate immune stimuli show a large degree of heritability (12, 29). We were curious to investigate whether there may be genetic variants associated with differential silencing of TNF release. Our initial finding that the most significantly associated SNP from a panel of over 48,000 SNPs was located in the promoter region for TNFR2, a receptor for the measured parameter (TNF release), was of high biological plausibility. We proceeded to demonstrate that this SNP, rs522807, marks a haplotype associated with increased basal expression of TNFR2 in our dataset and directly modulated allele-specific recruitment of a CRE binding protein. The observation that the basal expression of TNFR2 relates to secondary TNF release was replicated across the cohort, independently of genotype, with a highly significant correlation between expression and secondary response. By contrast, there was no association with the expression of TNFR1, suggesting a specific role for TNFR2 in the regulation of LPS-induced TNF release. This may reflect the differential signaling pathways of TNFR1 versus TNFR2, with TNFR2 specifically activated by membrane-bound TNF and showing relative insensitivity to free TNF (27). It is plausible that the early expression of membrane-bound TNF in response to LPS and subsequent signaling via TNFR2 has later effects on tolerance. This would also explain why the basal expression levels on TNFR2 seem to be of greatest importance in influencing subsequent later responses, with individuals expressing higher levels having consequentially reduced tolerance. We substantiated this finding by using mAbs specific for TNFR2 that enhance the affinity of the receptor for TNF (27). Pretreatment of cells with Ab has no stimulatory effect in its own right, demonstrating that TNFR2 activity only acts to amplify other responses in this experimental model. Moreover, at 2 h, cells pretreated with Ab show similar LPS-induced TNF release to those pretreated with isotype control. However by 6 h, a time when endotoxin tolerance has developed (20), there is a highly significant difference between isotype control-treated cells and those treated with TNFR2-specific Abs, indicating positive feedback of LPS-induced TNF release, acting through the receptor to enhance further TNF production. Using a purified monocyte subset, we were able to separately demonstrate that activity involving TNFR2 could modulate tolerance, with monocytes pretreated showing a significant increase in secondary TNF release compared with control cells.
Our data have highlighted the extent of individual variation in the endotoxin tolerance phenotype among healthy individuals and has implications for the tailored use of therapeutic interventions to modulate the inflammatory response. We have shown how expression quantitative trait mapping in primary human tissue can be used to identify functionally relevant polymorphisms for complex immunological responses. This unbiased approach allows the elucidation of pathway genes for which the role may have been previously unsuspected—in this case, the TNFR2 gene in endotoxin tolerance. This result has significant physiological implications for our understanding of the biological mechanisms regulating endotoxin tolerance and highlights the value of large-scale genotyping in the identification of functional variation in innate immune responses in man.
The authors have no financial conflicts of interest.
We thank all of the volunteers who participated in this study and members of the Knight laboratory for helpful discussion and suggestions.
This work was supported by the Wellcome Trust (Grants 074318 [to J.C.K.], 088891 [to B.P.F.], and 075491/Z/04 [to core facilities Wellcome Trust Centre for Human Genetics]).
The online version of this paper contains supplemental material.
Abbreviations used in this article:
- confidence interval
- cAMP response element
- linkage disequilibrium
- mi
- micro
- single nucleotide polymorphism
- TNFR-associated factor
- variable number tandem repeat.
- Received May 28, 2010.
- Accepted December 28, 2010.
- Copyright © 2011 by The American Association of Immunologists, Inc.


text: Shanghai Huaji Information Technology Co., Ltd. is a leading provider of exhibition technology service in China. It is headquartered in Shanghai Zhangjiang Hi-Tech Park, with branches in Beijing, Guangzhou and Shenzhen, and its business covers the whole country. Huaji focuses on the research and development and service of high-tech products in MICE industry, specializing in the broadband network deployment, the precise marketing of the exhibition big data, one-stop commericial live broadcast and other services. Up to now, Huaji has provided 3000+ technical services for 1000+ enterprise customers. Huaji adheres to the customer-centered service concept and relies on high-tech products and high-quality services to help more and more enterprise customers to achieve brand and commercial success.


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Management of mining quarrying and oreprocessing waste in the european union this project was completed mainly through the use of questionnaire sent to subcontractors in almost each country of the eu to assess this information and to extrapolate to the next twenty years this approach has been reinforced using publishedMore Details
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A quarry is a place where rocks sand or minerals are extracted from the surface of the earth a quarry is a type of mine called an openpit mine because it is open to the earths r type of mine a subsurface mine consists of underground tunnels or shafts the most common purpose of quarries is to extract stone for building materialsMore Details
Quarry byproducts are basically untried in field flowable fill applications experience is needed in the design placement and performance of flowable fill mixes containing different sources of quarry byproducts including some mixes with coal fly ashMore Details
Aggregates from waste marble may meet the huge demand for aggregates by the pavement construction industry the work aims at studying the use of waste marble fragments generated during the production of marble blocks and cutting processes as aggregates in the asphaltic mix design procedureMore Details
The montorfano and baveno granite ore bodies are located in the lake district vcone piedmont they were and are still quarried as dimension stones with a consequent production of a huge volume of waste in 1995 an italian company gmm spa decided to invest in the valorisation of granite quarry waste as a secondary raw materialMore Details
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text: Home / Shop / Toys and Games / Toys / DAY- DREAMIMAL SHARKIE – Backpack Keychain DAY- DREAMIMAL SHARKIE – Backpack Keychain$16.90 In stock Full description DAY- DREAMIMAL SHARKIE - Backpack Keychain quantity — OR — Add to cart Description Reviews (0) Description Hi, I am Sharkie! I am soft & confident and I'm the most adventurous of all the Dreamimals™. I love snuggling and sharing fun dreams. Plus, Whenever you have a negative thought, my specialty is inspiring your personal power to think positive. Dreamimals™ Sharkie Keychain Comes with Sharkie Keychain with Dream Pocket & 20 Dream Wishes. Machine Wash & Dry. 4″ wide Reviews There are no reviews yet. Be the first to review "DAY- DREAMIMAL SHARKIE – Backpack Keychain" Cancel replyYour rating * Rate… Perfect Good Average Not that bad Very poor Your review *Name * Email * Save my name, email, and website in this browser for the next time I comment. Current ye@r * Leave this field empty Related products Custom Coordinates Bracelet0 out of 5 $21.54 Select options Field Pack0 out of 5 $53.00 Add to cart Socksmith Men's Socktopus0 out of 5 $21.99 Select options


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Chase Bank offers free DoorDash food delivery to credit card holders: Here's how to get it
How hungry are you right now? Hungry enough that you would pay extra for food delivery?
Probably, but you may not have to if you own a Chase credit card. The financial giant just announced a partnership with DoorDash to offer free subscriptions to DashPass, the food-delivery company's subscription service. Specifically, the offer is available to people who own Chase Sapphire, Freedom, and Slate cards.
To take advantage of the offer, all you have to do is set an eligible Chase card as your default payment method in the DoorDash app and activate a DashPass membership by December 31, 2021, which means you have plenty of time. Once that's done, you'll get the following:
Note that neither offer is permanent—meaning once it's over, you will have to pay for DoorDash's "free" delivery service, and that currently costs $10 a month.
Teaming up with Chase is a smart move for the six-year-old DoorDash as it continues to eat up market share from competitors like GrubHub and Uber Eats in an overly crowded space. According to recent data from Second Measure, DoorDash had about 37% of the meal-delivery market in November.
If you're an eligible cardholder, you can activate your membership here. Evergreen PSA: Just remember to tip the delivery person!


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North Side News
August 27th - 31st
Week 4 of school
Have you reached out about something positive? It takes a village to raise a child, we are part of the village, but we must have parents working with us, make that positive connection! This will help parents to feel comfortable to reach out to you as well.
Coffee and texting/social media
How do you relax and de-stress?
Lay down, meditate, nap, deep breathing
How do you relax and de-stress?
Snuggling with your pet(s)
How do you relax and de-stress?
Monday Morning PD: Mindfulness - everyone's social and emotional well-being/PBIS
This Week: Aug. 27th - 30th
BOY Benchmarking - NWEA
Monday, August 27th:
PD: Mindfulness - Social and Emotional Well-Being/PBIS
8:30 a.m. - NWEA 4th gr. Reading
11:30-3:30 p.m. - Stephanie and Cory at Lawyer Session @ Impact
Tuesday, August 28th:
8:25 a.m. - NWEA 5th gr. Lang. Arts
8:30 a.m. - NWEA 3rd gr. Reading
3 p.m. - Maker/Tech Meeting with Chelsey in Makerspace room (Optional for specials teachers)
6 p.m. - Kindergarten Parent Meeting over Pilot Report Card (NS cafeteria)
Wednesday, August 29th:
8:25 a.m. - NWEA 5th gr. Reading
8:30 a.m. - NWEA 4th gr. Math
Lunchtime - Popsicle sold by the PTO
3 p.m. - Reading Committee Meeting (conference room)
Thursday, August 30th:
**Bus Evacuation Practice**
8:30 a.m. - NWEA 3rd gr. Lang, Arts
8:30 a.m. - NWEA 4th gr. Lang, Arts
12:40 p.m. - NWEA 5th gr. Math
3 p.m. - Leadership Meeting (Conference room)
Friday, August 31st:
NWEA Make-Ups
"Life is a dance. Mindfulness is witnessing that dance!" -Amit Ray-
Week of September 3rd - 7th
***Both special education and Title I LLI groups will be starting this week - remember they must stagger due to book usage so all groups will not start on Tuesday!Monday, September 3rd:
Labor Day - No School
Tuesday, September 4th:
11:55 a.m. - 5th grade (Davis room)
12:50 p.m. - 1st grade (Wible's room)
1:45 p.m. - 3rd grade (Whitney's room)
3 p.m. - Math Committee Meeting
Wednesday, September 5th:
11:55 a.m. - 4th grade (Maag's room)
12:50 p.m. - Kindergarten (Zollar's room)
1:45 p.m. - 2nd grade (Glasper's room)
3 p.m. - Leadership Meeting
6 p.m. - Board Meeting
Thursday, September 6th:
8:30 - 9:45 a.m. - Author Visit for grades 3-5
8:30 a.m. - Principal's Meeting (Stephanie)
3 p.m. - RTI/BIP Update - L.H. (Jessica, Stephanie, Cory, Jamie, Amanda H., Laura P., Anita)
Friday, September 7th:
3 p.m. - Meeting on N.M. (Laura P., Laura S., Stephanie, Erin)


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Location Location Location, Grandezza's highly desirable Grande Estates gated community welcomes you to this gorgeous 3275 sqft home with 4 bedrooms, 5 baths, den/office and large 3 car garage. This stylish design homes features an open floor plan with luxury finishes such as crown molding throughout the home, marble and wood flooring, 14ft/16ft ceilings, large master suite with 2 custom walk-in closets, walk-in shower with 3 shower heads, whirlpool jacuzzi tub and his & hers private bathrooms! The gourmet kitchen features Viking gas cooktop, Subzero, GE Profile and Bosch appliances. The laundry room has an additional refrigerator, LG steam washer and 2 LG TROMM dryers with lots of storage and utility sink. The lavish outdoor area is perfect for entertaining and privacy with Preserve views, heated lap pool and spa, granite counter bar area with mini refrigerator, exterior and interior surround sound, hurricane impact door and windows, new a/c and new roof (2019) make this a must see home. The club of Grandezza features championship golf, tennis and social membership levels for an additional fee. Ask about our Video tour
Below Ground, Heated Gas, Lap Pool, Screened, Self Cleaning
Basketball, BBQ - Picnic, Bike And Jog Path, Bike Storage, Bocce Court, Business Center, Cabana, Clubhouse, Community Pool, Community Room, Community Spa/Hot tub, Exercise Room, Fitness Center Attended, Golf Course, Hobby Room, Internet Access, Library, Private Membership, Putting Green, Restaurant, Sidewalk, Streetlight, Tennis Court, Underground Utility
2+ Spaces, Driveway Paved, Golf Cart
Bar, Built-In Cabinets, Cable Prewire, Cathedral Ceiling, Closet Cabinets, Foyer, French Doors, High Speed Available, Laundry Tub, Pantry, Pull Down Stairs, Smoke Detectors, Surround Sound Wired, Tray Ceiling, Walk-In Closet, Window Coverings
Central Electric, Gas - Propane
Ceiling Fans, Central Electric, Gas - Natural, Gas - Propane, Humidistat
Built In Grill, Outdoor Kitchen, Outdoor Shower, Sprinkler Auto
Concrete Block, Poured Concrete
The source of this real property information is the copyrighted and proprietary database compilation of the Southwest Florida MLS organizations Copyright 2020 Southwest Florida MLS organizations.. All rights reserved. The accuracy of this information is not warranted or guaranteed. This information should be independently verified if any person intends to engage in a transaction in reliance upon it.
The data relating to real estate for sale on this Website come in part from the Broker Reciprocity Program (BR Program) of M.L.S. of Naples, Inc. Properties listed with brokerage firms other than Equity Realty are marked with the BR Program Icon or the BR House Icon and detailed information about them includes the name of the Listing Brokers. The properties displayed may not be all the properties available through the BR Program.


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Intro – Erie Federal Credit Union Reviews & Ranking
AdvisoryHQ recently published its review and ranking of the top credit unions in Pennsylvania, a list that included Erie Federal Credit Union.
Below we have highlighted some of the many reasons Erie Federal Credit Union was selected as one of the best credit unions in Pennsylvania.
Click here for a detailed review of AdvisoryHQ's selection methodology: AdvisoryHQ's Methodology for Selecting Top Banks and Credit Unions.
Erie Federal Credit Union Review
Erie Federal Credit Union is a community-chartered credit union that has been honoring its mission to responsibly deliver a wide range of financial services to meet the needs of its membership for the past eighty years. They continue to strive to serve their members with spirit, compassion, and heart.
This Pennsylvania credit union has grown tremendously since first being formed in 1936 by a group of local Erie educators, who operated out of a board member's house. Today Erie Federal Credit Union boasts nine branch locations, $458 million in assets, and more than 53,000 members.
Image Source: Erie Federal Credit Union
Key Factors Leading Us to Rank This Firm as One of This Year's Top Credit Union Firms
Upon completing our detailed review, Erie Federal Credit Union was included in AdvisoryHQ's ranking of this year's best credit unions based on the following factors.
Erie Federal Credit Union Review: Flexible Checking Account Options
Erie Federal Credit Union offers several flexible checking account options to allow members to choose which options they want and which they don't. Checking accounts at this credit union in PA offer many of the same features as big-name banks without the hidden and complicated maintenance fees.
Whether you like your checking accounts kept simple and sweet or wish to earn higher than average monthly dividends on your funds, this top Pennsylvania credit union has a checking account for you.
Checking account options are:
- Free Checking: no surprises here, free checking with no minimum balance requirements and a free box of checks with direct deposit
- Premium Checking: interest-earning checking account with no balance requirements, $2 monthly maintenance fee, and a free box of checks with direct deposit
- Dividend Plus Checking: interest-earning checking account with no minimum balance requirement but requires a minimum average daily balance of $500 to earn a higher dividend rate, $2 monthly fee can be waived if requirements are met, a free box of checks with direct deposit.
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Erie Federal Credit Union Review: Business Deposit Accounts
From checking accounts to savings and money market accounts, Erie Federal Credit Union offers a variety of business deposit accounts to help businesses succeed and grow their bottom line.
Business deposit accounts available at this credit union in Pennsylvania are:
- Business Checking: an interest-earning business checking account for businesses with high transaction volumes
- Free Business Checking: a low-cost, non-interest-earning checking account solution for businesses with low transaction volumes
- Business Savings: a great way for businesses to begin building excess funds
- Money Market and Share Certificates: helps businesses earn higher dividends on long-term savings
Erie Federal Credit Union Review: Mortgage Realtor Rebate Program
Erie Federal Credit Union is a Pennsylvania credit union that strives to offer more with their home loans than the average financial institution does. The cost of purchasing a home can be staggering, a fact that Erie FCU understands well. For this reason, they offer members their Realtor Rebate Program.
The Realtor Rebate Program allows members to earn up to 25% off their realtor's commission on the purchase or sale of their home. Members simply select an approved real estate agent from Erie FCU's website and register for the Realtor Rebate Program. It's that simple.
Image Source: Erie Federal Credit Union
The Realtor Rebate Program offered by this top credit union in PA can potentially save members hundreds or even thousands of dollars on the sale or purchase of a home. But there are many other great benefits to choosing a mortgage from Erie FCU:
- Free mortgage pre-approvals
- Competitive interest rates
- Lower mortgage fees
- Locate an experienced real estate agent
In addition to the above Erie Federal Credit Union review, you can click on any of the links below to browse exclusive reviews of AdvisoryHQ's top rated banking firms & credit unions:
Top Rated Banks
Top Banking Firms
Review of Top Mortgage Firms
Bank Reviews
AdvisoryHQ (AHQ) Disclaimer:
Reasonable efforts have been made by AdvisoryHQ to present accurate information, however all info is presented without warranty. Review AdvisoryHQ's Terms for details. Also review each firm's site for the most updated data, rates and info.
Note: Firms and products, including the one(s) reviewed above, may be AdvisoryHQ's affiliates. Click to view AdvisoryHQ's advertiser disclosures.


text: I have recently completed my CBT and MOD 2 and would like to thank Paul for making it a pleasurable experience. From start to finish Paul was welcoming, patient and as a teacher myself I fully appreciate his excellent teaching skills. Having received a poor experience at one of the Islands other training centers I can say without doubt that Pitstop offer an excellent service. One of sons has a also trained with Pitstop and my youngest son who has just turned 16 will be training with him in the near future.Allowing my children to train with Pitstop is the biggest complement I can offer.


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DJ & Producer Olivia Jaxx Dies
Such sad news that the lovely and talented DJ & Producer Olivia Jaxx has Died.
Olivia was a growing talent on the London scene with residencies at Casita and was due to play on 14th March for their show at E1 London. She was also a regular for Ritual, Mas Vida, Edge Events and Ministry of Sound.
We'd been a fan of hers for a couple of years and she'd been growing into a great young talent. We'd been watching her progress after spotting her on a room 3 line-up for Ministry of Sound and she'd always been supportive of our Shanghaied label. So it's very upsetting to hear her passing. 2020 had been looking a good year for her with her announcing her first music (below) with Taylor Crane and big bookings at We Are FSTVL.
Other than what has been put on friends social media, no other information is available about her death.
Tributes from her friends have appeared on social media. Hot Creations DJ & Producer WaFF has said "I can't believe I am even writing this. I'm in complete shock. Absolutely devastated. Right know we don't know the circumstances of your passing. 😢😢 But Im so sorry Olivia. I will forever remember you and love you. Your were a big part of my life. We have shared so much together, and became one of my best friends. I will never forget all the laughs we had, and all the times killing each other on Halo 🤣!"
Dale Hart from Hart & Neenan & Metafloor said: As some people are aware – Olivia Jax (Sacha) and I were really close friends and music first connected us together. The last 2yrs Olivia became one of my closest mates and to wake up today and hear shes been taken like this has actually destroyed me so much… After speaking to her on Saturday night she was in high spirits and we were due to catch up this week. A lot of people saw the happy side to Olivia – who would usually look flawless or turn up to a gig and blow the crowd away for her incredible DJ ability. Watching her develop as a Female artist professionally and remain so humble during the process was something I was so fucking proud off.
Our condolences to the family & friends and she now rest in peace.
Please can you help the family raise money for the funeral, by just giving here.


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Direct Selling: Picture Perfect
Close your eyes and think of a Pottery Barn catalog. Or Lands' End. Or J. Jill. Can you picture them? Chances are, you have a clear and distinct image in mind. You know what the photography looks like because all of these brands have established a certain style and "look" that is unique to them. They've achieved this over time through consistent execution of their defined styles.
Pottery Barn, for instance, always shows a warm and inviting room consisting of the various pieces of furniture and accessories sold in the photo spread. It allows you to picture yourself in that perfect environment. Lands' End, on the other hand, focuses on the details of its garments. Quality is key, so by showing unique fabrics or the stitching on a button up close, the cataloger can emphasize the attention to detail given to all its products. J. Jill shows off its clothing on models, typically outside in a natural setting, portraying a casual and comfortable lifestyle. All of these creative decisions are made with each catalog's specific brand in mind. The fact that they maintain consistency over time is the secret to their success.
Branding is a hot topic in business today. In a competitive and saturated environment, it's essential that you set yourself apart from the competition by creating a memorable and relevant brand. One step to achieving this goal is by using photography as a differentiator.
Give Your Products a Face
Branding is much more than good photography. But a distinctive, recognizable photographic style can go a long way to helping you establish a strong brand. Brands should be "experiential"—meaning customers should be able to experience your brand on many different levels using all of their senses. Consumers have an advantage when they're shopping at a retail location because they can put all their senses to use and experience products. But when shopping through the direct channel, consumers have to rely far more on sight. So as direct marketers, you're challenged to visually communicate everything about your brand. You need to harness the power of photography to help you stand out from the crowd.
Ask yourself this—if you were to cover up your name or logo on the pages of your catalog, Web site or direct mail piece, would your audience still know it was you who sent it? To further your brand, your photography should accomplish the following five goals:
1. Create desire. You want people to buy your products, so you must show them off in the most desirable way. The right lighting and props can create the right mood and make your products appear more attractive. Just don't overdo it. Lighting that is too dramatic or props that are distracting will take focus away from what you're selling.
2. Be consistent. You will get tired of your photography long before your customers do. That's a fact. Resist the urge to experiment with different styles or looks. Consistency is the key to building a brand, and photography is no exception. Just imagine if Pottery Barn decided to alter its photographic style on a whim every now and then. It might keep its art directors creatively challenged, but it would only create confusion among its loyal customers. Find a style that is uniquely yours and stick with it.
3. Be distinctive. So many companies want to imitate the market leader. They assume that if it works for the leader then it should work for them. All that does is help the leader by reinforcing its brand. If you want to help yourself, take this advice—when the market leader zigs, you should zag.
4. Reflect the personality of the brand. This allows you to put a face on what is otherwise an inanimate object, increasing the likelihood that the audience will identify and connect with the product on a personal and emotional level. If your brand is fun and playful, your photography should be fun and playful. If your company's image is upscale and refined, your photography should reflect that. Think of it in terms of a real person. When you think of James Bond, you picture him wearing a tuxedo. Who or what does your brand look like?
5. Show your products clearly and accurately. Make sure the product always is the hero of your photography, not the props or the styling. Even though you want to build a desirable brand and communicate it through photography, the ultimate goal is to sell more products. Since the customer can't pick it up and touch it, smell it, or taste it, the important aspects of the product always must be in focus and easy to see.
Attaining these goals will help you achieve the ultimate objective, which is an increase in sales.
Before You Snap the Shutter …
To get the desired results from your photography, you have to do some prep work. Here are some pointers to consider when preparing for a photo shoot:
Always hire a professional photographer, even though it might be tempting to save some money and shoot the photos yourself. The fact that you own a car doesn't qualify you to be a NASCAR driver. Likewise, owning a digital camera doesn't make you a good photographer.
Have a plan and stick with it. Clearly understand what makes your brand different, and decide in advance how you will communicate this concept with your photography. Meet with your photographer, art director and stylist first to agree on how to execute each photo. If everyone is on the same page before the photo shoot starts, everything will run more smoothly on set.
Work from an existing layout. Whenever possible, have the layout of your catalog or direct mail piece finished before you begin your photo shoot. This will help you decide which images are heroes, which direction the product needs to face and how multiple photos work together on a spread. You may consider having sketches of each photo on hand to indicate angle and props.
Dramatic, distinctive photography can go a long way in setting your brand apart in a sea of sameness. It can add stopping power to the cover of a catalog; instantly welcome customers to the homepage of a Web site; and add a sense of familiarity to a direct mail piece. And above all, it can help you increase sales. Picture that.
Brent Niemuth is creative director and brand evangelist at J. Schmid & Associates, Mission, Kan. You can reach him by e-mail at email@example.com.


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What is the difference between Financed and Hard Money in a Short Sale
There are common misconceptions regarding the terms, "Financed vs Hard Money" when purchasing a short sale in Massachusetts or any state.
Please keep in mind if you're an investor of a short sale when you are putting an offer in and you know you're going to get hard money what that truly means is you are financing the offer.
Please don't put in your purchase contract that it's a cash offer. It isn't. You can put financed not based on appraisal, but don't put in that it's a cash offer.
The seller's lender does not care if you're financed, if you're hard money, or if you're cash. The seller's lender on a short sale is looking for their bottom-line net. If their bottom-line net is going to net them more than what they would have to go through to put that property through foreclosure chances are your short sale will be approved.
The issue is hard money is actually still having to FINANCE a property. So, unless you truly have the cash sitting in an account, make sure when you put your offer in on your short sale you are letting your agent know that it's a financed offer, with only one contingency or no contingencies.
For Realtors, Attorneys, Title companies, etc interested in our services please go to http://massachusettslossmitigation.com/
For homeowners needing assistance with avoiding foreclosure please go to http://shortsalemitigation.net/
2500 Main Street,
Tewksbury, MA 01876


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June 15 – Reece Goforth of Springfield visited family and friends in Ava on Monday.
Macee Breeding and Donna Dodson came by on Monday.
Bryse Dodson worked in my yard on Tuesday. Donna was also here.
Donna and Macee came by on Wednesday and Thursday.
Megan Goforth attended a friend's graduation party on Friday. She and Bradley Williams of Branson attended graduation Friday night in Ava. Keith and Melanie Breeding, Quin and Macee also went.
Mike Dodson visited Cassandra Cornett and Ruger, David and Donna, Keith and Melanie, and me this week. Bryse helped him several days this week.
Donna and Macee came by on Friday. Macee has been painting her bedroom.
On Saturday, Donna and I went to lunch then got groceries, then later went to dinner at a local restaurant. They had a good singer there. Diane Davis joined us for supper. She and Butch had been to the creek with Amy and Ronnie Croney and family.
Keith and Melanie, Megan, Quin, and Macee attended Regan Koop's graduation party Saturday afternoon.
Bradley Williams and Megan Goforth visited Dale and Glenda Evans and Charlie Saturday evening.
Quin and Macee and Cass and Mike all visited David and Donna on Saturday.
David Dodson has been sick over the weekend.
Bryse Dodson mowed my yard Sunday.
Reece and August of Springfield spent Sunday night with David and Donna.


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Alibaba.com offers 20,566 building boilers products. About 39% of these are boilers, 1% are heat exchanger, and 1% are other machinery amp; industry equipment. A wide variety of building boilers options are available to you, such as once through, natural circulation.
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|Form when limestone is dissolved by acidic waters
|caves and sinkholes
|Examples of soft rocks that are affected by acidic waters:
|marble and limestone
|rapid movement of a large amount of rock and soil
|What is erosion?
|movement of fragments of rock and soil
|What are four forces of erosion?
|water, wind, ice, and gravity
|What factors speed up chemical weathering?
|warm temperatures, abundant rainfall, and thick vegetation
|What is the other name for mechanical weathering?
|physical weathering
|There are two types of weathering, what are they?
|chemical and mechanical
|What factors speed up physical weathering?
|cool climates
|What is frost wedging?
|Repeated freezing and thawing of water in cracks that physically breaks apart rock.
|What is weathering?
|breaking down or changing materials
|Why does an object with a larger surface area break down faster?
|A larger area is exposed to weathering forces.
|Name the three different kinds of rock.
|metamorphic, igneous, and sedimentary
|How does igneous rock form?
|cooling and crystallization of magma
|How does sedimentary rock form?
|compacting and cementing of sediments
|How does metamorphic rock form?
|heat and pressure deep in the earth
|How does magma form?
|melting of solid rock
|How does sediment form?
|weathering of rock
|What is transpiration?
|Water evaporating from plants
|What is infiltration?
|Water soaking into the soil.
|What types of water movement are due to gravity?
|assimilation, runoff, infiltration, precipitation
|How does the rate of precipitation affect infiltration?
|If rain falls too fast then the soil becomes saturated and water ca no longer infiltrate it, then it becomes runoff.
|Does water use energy or release energy to go from solid to liquid to gas?
|Does water become more or less mobile going from solid, to liquid, to gas?
|Why does it require more energy to change water from solid, to liquid, to gas?
|because the water molecules are becoming more mobile.
|What slows runoff and increases infiltration?


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Yes, you read that right, gift cards, paypal cash and rewards coming your way for free.
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Scientology Network's MEET A SCIENTOLOGIST, the weekly series spotlighting the everyday lives of Scientologists from around the world and all walks of life, announces an episode featuring the award-winning singer-songwriter Michele Henderson.
MEET A SCIENTOLOGIST airs Tuesdays at 8 p.m. ET/PT on Scientology Network.
Michele Henderson is an award-winning
singer-songwriter hailing from the island of Dominica in the West Indies.
Drawing on her passion to inspire others with her art, she's performed for
international audiences and penned the song "Celebrating CARICOM (Caribbean
Community)," which would go on to become the official anthem of Caribbean
Michele Henderson was born in the Caribbean island nation of Dominica. From the age of two she was already developing and honing her musical skills by singing in the church where her father worked as the musical director. Her parents encouraged and nurtured her obvious musical talents throughout her upbringing, enrolling her in music school where she studied flute and music theory and teaching her the craft of songwriting. At the age of 15, Michele entered and took first place in a songwriting contest. The experience served as a confirmation of her talent and gave her the confidence to pursue music as her livelihood. Michele married former band mate, Roland Delsol Jr., who produced her first solo album, self-titled, Michele Henderson. Developing her repertoire in a number of genres, including jazz, calypso, reggae, R&B, soul and Zouk (a musical style developed in the islands of Guadeloupe and Martinique), she racked up hit songs and star turns at music festivals throughout the world. She's shared the stage with such musical virtuosos as Chick Corea and appeared on the bill with Green Day and the Foo Fighters. Michele's popularity soared throughout the Caribbean and she was appointed an Official Goodwill Ambassador to the Commonwealth of Dominica.
The Scientology Network debuted on March 12, 2018. Since launching, the Scientology Network has been viewed in 240 countries and territories worldwide in 17 languages. Satisfying the curiosity of people about Scientology, the network takes viewers across six continents, spotlighting the everyday lives of Scientologists; showing the Church as a global organization; and presenting its social betterment programs that have touched the lives of millions worldwide. The network also showcases documentaries by Independent filmmakers who represent a cross section of cultures and faiths, but share a common purpose of uplifting communities.
Broadcast from Scientology Media Productions, the Church's global media center in Los Angeles, the Scientology Network can be streamed at scientology.tv and is available through satellite television, mobile apps and via the Roku, Amazon Fire and Apple TV platforms.
Media Relations
(323) 960-3500
Stay tuned to the Scientology Network.
In its very first year of entering the competition, Scientology Media Productions announces eight wins at the 2020 dotCOMM Awards, the annual international competition honoring excellence in web creativity and digital communication.
The Ukrainian psychiatric industry rakes in well over $2 billion each year by stripping its citizens of their legal rights, subjecting them to abuse and human rights violations in institutions across the country.
Two decades after the end of a civil war that claimed 200,000 lives, the people of Guatemala still struggle with the human rights abuses committed by their military.


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The closure of the Hungarian border will force migrants to go through the territory of Croatia. And there on their way can meet real minefields.
The mined areas remained in Croatia after the Balkan wars of the 1990s. Despite the fact that cleaning up of territories has been underway for several decades, there are still dangerous zones in Croatia. Even more in Bosnia and Herzegovina. But it is in Croatia that the main concentration of mined areas falls on areas bordering on Hungary
The Croatian edition of the Diary reports that the local police have already prepared themselves for refugee flows.
"Croatia is ready for a wave of migrants. The police will register each person, there will not be a mass pass, as in some other countries, "the newspaper said.
Meanwhile, the situation in Europe, including in Hungary, is heating up. Two days ago, refugees broke a four-meter fence, which was installed at the border control. Budapest has introduced a number of amendments, which come into force on 15 September. They tighten legislation on migration and border protection. The innovations envisage the army's participation in the protection of state borders, the prosecution of those who illegally cross the border and the accelerated procedure for examining asylum requests.
Other countries also tighten border controls. Germany first suspended the Schengen Agreement. Between Germany and Austria, for the first time in two decades, a border pillar appeared. Political scientists already argue that the situation in Europe can lead to the collapse of Schengen


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Shady Sayed is an Arab Weekly
contributor in Egypt.
Cairo - Three years ago, Sultan Mohamed made a "transforming" visit to Germany.
CAIRO -Standing in the parliamentary elections and winning a seat was a dream long cherished by
CAIRO - With polls due to open on October 17th, these are busy days for 48-year-old Egyptian call
Cairo - The long Nile cruise from Cairo to the southernmost province of Aswan has been a dream j
CAIRO - Egypt's new government, sworn in after the previous administration resigned in a corrupti


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Meet me in Nashville!
January 22, 2020
I'm finally leaving the state of Michigan for some reason other than school! The conference hiatus has ended!
I will be appearing at Booklovers Con from March 18 to 22 in Nashville, Tn.
I will be signing multiple titles at the BookBash signing at 9:30 am, March 21. My other events are
9:30pm - 11:00pm, March 18
Governor's Chambers C
-Paranormal Romancelandia Feud
3:30 - 5 pm, March 19
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12:00 - 1:00pm, March 20
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- Ethnic group. Closely related to the Adyghe, Kabardins or Kabards are the easternmost and the largest subgroup of the Circassian people, which also includes the Adyghe, Cherkess, Abazins, and Abkhaz. They speak a language from the Northwest Caucasian family which also includes Adyghe. The Kabardins number more than 500,000 and are the titular majority in the Republic of Kabardino-Balkariya, which they share with the Turkic Balkars. Historically, the Kabardins have been the most pro-Russian of the Caucasian mountaineers. Unlike their western Circassian counterparts, their feudal elite were co-opted by tsarist Russia. Most Kabardins are Sunni Muslims, converting to Islam from Christianity under Ottoman influence; however, their identification with Islam remains comparatively low, even by Caucasian standards. Some noble families converted to Russian Orthodoxy after the Romanov conquest and still retain their Christian faith. Tsarist and Soviet policies of favoritism toward the Kabardins color interethnic relations in the republic, especially given the Stalin-era deportation of the Balkars. Economically and politically, the Kabardins are the dominant force in post-Soviet Kabardino-Balkariya. Unlike the Balkars, the Kabardins possess a well-developed sense of national identity that is reinforced by traditional codes of loyalty and social hierarchy; as such, they are not generally attracted to pan-Islamic ideals. However, this has not prevented some Kabardins from joining Islamist terrorist groups like Yarmuk.
Historical Dictionary of the Russian Federation. Robert A. Saunders and Vlad Strukov. 2010.
Look at other dictionaries:
Circassian nationalism — Circassia. Alexander Keith Johnston, 1861 Circassian nationalism[1] is the desire among Circassians to establish (or rather, revive) an … Wikipedia
History of Chechnya — The History of Chechnya refers to the history of Chechens, Chechnya, and the land of Ichkeria. Chechen society has traditionally been organized around many autonomous local clans, called taips. The traditional Chechen saying goes that the members … Wikipedia
Yarmuk Jamaat — Infobox Military Unit unit name= Yarmuk Jamaat caption= dates= 2002 Present allegiance= type= Infantry size= battles= Second Chechen War commander1= Muslim AtayevKIA Anzor Astemirov commander1 label= Commanders Yarmuk Jamaat is a militant… … Wikipedia
North Caucasian peoples — is a term used to describe a peoples speaking North Caucasian languages.PeoplesNorth Caucasian peoples are: *Northwest Caucasian peoples: **Abkhazians **Abazins **Adyghe **Cherkess **Kabardins **Ubykh *Nakh peoples: **Bats **Kists **Chechens… … Wikipedia
Kabardie — La Kabardie (Кабарда en russe, Къэбэрдей en tcherkesse), autrefois Tcherkessie de Piatigorsk est une région historique caucasienne en fédération de Russie. Elle doit son nom au peuple des Kabardes ou Kabardins, proches des Adyguéens. Cette région … Wikipédia en Français
Balkars — Ethnic group. The Balkars are a Turkic ethnic group of some 108,000 inhabiting the North Caucasus. They are sometimes referred to as Malkars, though many self identify as Taulu ("mountaineers" in Karachay Balkar). They are one of two titular… … Historical Dictionary of the Russian Federation
Kabardino-Balkariya — An ethnic republic of the Russian Federation. Initially part of the Mountain Peoples' Autonomous Republic of Soviet Russia, the region was organized into the KabardinBalkar Autonomous Oblast in the early 1920s, becoming an Autonomous Soviet… … Historical Dictionary of the Russian Federation
Caucasus — The Caucasus ( also referred to as North Caucasus ) is a geopolitical region located between Europe, Asia Middle East. It is one of the most linguistically and culturally diverse regions on Earth, home to Europe s highest mountains ( Mount Elbrus … Wikipedia
South Ossetia — Republic of South Ossetia Республикæ Хуссар Ирыстон / Respublikæ Xussar Iryston (Ossetic) სამხრეთი ოსეთი / Samkhret Oseti (Georgian) Республика Южная Осетия / Respublika Yuzhnaya Osetiya (Russian) … Wikipedia
Chechen people — Chechens (Noxçiy) … Wikipedia


text: Along with real estate marketing and design, I am also a licensed California Real Estate Agent and have worked on more than 400 real estate transactions. I offer Transaction Coordinator services which includes managing the paperwork from the beginning of a transaction to the close. My fee is paid upon closing and I can ensure that you are DRE compliant with all your paperwork. For more information please email me at: