Breast cancer is a heterogeneous disease for which prognosis and treatment strategies are largely governed by the receptor status (estrogen, progesterone and Her2) of the tumor cells. Gene expression profiling of whole breast tumors further stratifies breast cancer into several molecular subtypes which also co-segregate with the receptor status of the tumor cells. We postulated that cancer associated fibroblasts (CAFs) within the tumor stroma may exhibit subtype specific gene expression profiles and thus contribute to the biology of the disease in a subtype specific manner. Several studies have reported gene expression profile differences between CAFs and normal breast fibroblasts but in none of these studies were the results stratified based on tumor subtypes.
To address whether gene expression in breast cancer associated fibroblasts varies between breast cancer subtypes, we compared the gene expression profiles of early passage primary CAFs isolated from twenty human breast cancer samples representing three main subtypes; seven ER+, seven triple negative (TNBC) and six Her2+.
We observed significant expression differences between CAFs derived from Her2+ breast cancer and CAFs from TNBC and ER + cancers, particularly in pathways associated with cytoskeleton and integrin signaling. In the case of Her2+ breast cancer, the signaling pathways found to be selectively up regulated in CAFs likely contribute to the enhanced migration of breast cancer cells in transwell assays and may contribute to the unfavorable prognosis of Her2+ breast cancer.
These data demonstrate that in addition to the distinct molecular profiles that characterize the neoplastic cells, CAF gene expression is also differentially regulated in distinct subtypes of breast cancer.
Gene expression profiling of whole breast tumors has stratified breast cancer into several molecular subtypes that largely correlate with the expression status of three receptors in the tumor cells, namely estrogen (ER), progesterone (PR), and Her2-neu (Her2) [1, 2]. The most common breast cancer subtype expresses either ER or PR but lacks Her2 expression. Breast cancers that do not express any of the 3 receptors, known as triple negative breast cancer (TNBC), and those that express Her2 (Her2+) are less common, comprising approximately 15% and 25% of all breast cancers respectively. Her2+ and TNBC have less favorable prognosis compared to ER + cancers [3, 4]. How cancer cells acquire a specific molecular phenotype is uncertain. It has been postulated recently that the tumor stroma and the cancer cells may co-evolve to support the selection or enrichment of a specific cancer subtype .
Much of the earlier gene expression profile analyses of breast cancer were performed using RNA extracted from tumor samples comprised of at least 50% of tumor cells, with the tumor stromal cells being a minor but important component. As tumor cell survival and tumor progression are dependent on the tumor microenvironment, elucidating the symbiotic relationship between neoplastic cells and stromal cells is crucial to further our understanding of the pathogenesis of the disease [5–8]. This interdependency is reinforced by the recent identification of a stroma-derived gene signature that correlates with prognosis suggesting that the tumor stroma contributes significantly to the invasive and metastatic potential of tumor cells . A unique breast cancer stroma signature has also been observed in women of African American descent compared to European American descent , while a stromal gene signature has been reported to predict response to chemotherapy . These observations support the suggestion that intrinsic heterogeneities between the tumor stroma may correlate with patient-specific characteristics, prognosis, therapeutic response, and, perhaps, tumor subtypes. However, breast cancer subtype-specific differences have not yet been reported for the tumor stromal cells even though multiple studies have shown that the gene expression profiles of breast cancer associated fibroblasts (CAFs) are distinctly different from their normal counterparts. None of these prior studies had stratified their results based on tumor subtypes [12–16].
In this study, we isolated CAFs from twenty primary breast cancer samples representing three main subtypes (ER + (n = 7), TNBC (n = 7), Her2+ (n = 6)) and performed gene expression profile analyses on RNA isolated from these early passage CAFs. Subtype-specific gene expression profile differences were observed that distinguished CAFs derived from Her2+ cancers and TNBC and ER + cancers. Several genes, e.g. ITGA3, ITGA5, CFL1, and RHOA, that were found to be selectively up regulated in CAFs derived from Her2+ but not ER + or TNBC breast cancers are known to be involved with pathways associated with integrin and RhoA signaling suggesting that CAFs may contribute to the invasiveness of Her2+ breast cancer . Migration of breast cancer cells,T47D, was significantly enhanced by CAFs derived from Her2+ breast cancer compared with ER + or TNBC. Our findings suggest that CAFs might contribute to the biology of the disease in a subtype-specific manner. Our findings are also consistent with the recently proposed tumor-stroma co-evolution hypothesis .
Characterization of CAFs from breast cancer subtypes by morphology (light microscopy) and flow cytometry analysis. Top panel, 20x magnification light microscopy pictographs of a) ER+; b) TNBC and c) Her2+ breast cancer derived CAFs; Lower panel, histograms (dark solid line) depicting CAFs staining for (left to right): EpCAM, FAP, CD45, and CD31; light grey lines depict histogram of CAFs staining with isotype control antibodies.
Women with primary operable breast cancer undergoing breast surgery at the Hospital of the University of Pennsylvania were asked to participate in our tissue banking protocol approved by the institutional review board. Informed consent was obtained from all participants. Our study cohort included 20 women diagnosed with breast cancer between 2008 and 2011. Breast tumors were stratified into three subgroups according to receptor expression determined by immunohistochemistry (IHC) as described previously : 1) ER + denotes breast cancer which expresses either ER or PR and lacks Her2 expression (n = 7); 2) TNBC denotes breast cancer that lacks expression of ER, PR, and Her2 (n = 7); and 3) Her2+ group (n = 6) denotes breast cancer which expresses Her2 as determined by IHC and/or fluorescence in situ hybridization with (n = 1) or without expression of ER or PR (n = 5). All data collection and analyses were adherent to Institutional Review Board approved protocols. Clinical characteristics, including age at diagnosis, race, histology, tumor size, tumor grade, and number of involved (+) axilla nodes were compared. Pair-wise comparison was done using two-tail t-test for age and tumor size, and Fisher's exact test for race (Caucasian vs. African-American), histology, tumor grade (II vs. III) and number of (+) axilla nodes (none vs. one or more).
After our surgical pathologists completed gross examination and inking of the tumor specimen, fresh tumor tissue was taken from the center of the tumor without interfering with margin assessment as determined by the pathologists. The tissues were stored in ice cold medium DMEM/F12 supplemented with 10% fetal bovine serum (FBS), penicillin and streptomycin. The fresh tumor tissue was kept on ice at 4°C until ready for processing within 6 hours from the excision time. If the tumor tissue weighed less than 0.5 gram (n = 5) (TB160 – TB165), the tissue was mechanically dissociated by mincing with scalpel and scissors to 1–2 mm3 in a 10 cm tissue culture plate. Fibroblast growth medium (DMEM supplemented with 10% FBS penicillin and streptomycin) was then added. After several days, outgrowth of spindle shaped cells was observed. Tissue debris and non-adherent cells were removed and medium changed between day 2–4. For tissues (n = 14) weighing more than 0.5 gram (TB71 - TB148) the tissue was minced as described above and then enzymatically dissociated in tissue digestion buffer containing collagenase I (Worthington), hyaluronidase (Sigma), Collagenase IV (Worthington) at 1 mg/ml of each enzyme in DMEM/F12 medium in a volume of 1:5 ratio of tumor to buffer (wt/vol) on a gyrating platform at 37o C for 30 min. The digestion was quenched by addition of fibroblast growth medium and filtered through a 70 μm cell strainer. Cells were pelleted at 1500 rpm for 10 min. Tissue debris and non-adherent cells were removed during medium change between day 2 or 4. By 10 – 14 days, near confluent adherent spindle shaped cells were harvested using 0.25% trypsin in versene, washed and replated in fresh fibroblast growth medium. Medium was changed every 4 – 7 days. CAFs from early passages (passage 2–3) were harvested and the cell pellet was stored in RNA later (Applied Biosystems) at −80°C until RNA was isolated.
RNA purification was carried out using TRI Reagent® (Molecular Research Center) according to manufacturer's recommendations. RNA quality was determined using the Bioanalyzer (Agilent). Only samples with RIN numbers > 7.5 were used for further studies. Equal amounts (400 ng) of total RNA was amplified as recommended by Illumina and hybridized to the HumanHT-12 v4 human whole genome bead arrays. Illumina BeadStudio v.3.0 software was used to export expression levels and detect p-values for each probe of each sample. Quality control of each array was performed using median Spearman correlation computed against all other arrays. Arrays whose median correlation differed from the global correlation by more than 8 absolute deviations were marked as outliers and not used for further analysis (resulting in the removal of one TNBC sample, TB147 (Table 1)). The remaining 19 arrays were then quantile-normalized between each other and filtered to remove non-informative probes (probes with a detection p-value > 0.05 in all samples). Between-batch normalization was performed using Distance Weighted Discrimination (DWD) approach  using 4 samples replicated in the 2 microarray batches. Average expression between replicates was used for data analysis. The data was submitted to GEO database (http://www.ncbi.nlm.nih.gov/geo/) and available by using accession number GSE37614.
List of samples divided into two batches (b1 and b2) including two samples from each subtype as an independent validation (testing) set as indicated.
EpCAM: PE anti-human CD326 clone 9C4 (Biolegend) used at 1ug/ml; PE-F19: mouse anti-human FAPα monoclonal antibody (clone F19), used at 1/10 dilution, was purified from serum-free hybridoma supernatant as described [20, 21]; CD45: APC mouse anti-human CD45 (BD Pharmingen) used at 20ul/test according to manufacturer's recommendation; CD31: APC anti-human CD31 clone WM59 (eBioscience) used at 5ug/ml.
We randomly selected two samples from each Her2+, ER + and TNBC subtype as an independent validation set (testing set, Table 1). One sample which was unique in its subtype classification in that the CAF was derived from a Her2 + and ER + breast cancer (TB148, Additional file 1: Table S1) was also added to the testing set in order to show how it would be classified based only on its gene expression profile. The training set used to select the genes that distinguish the 3 CAF subtypes included 3 Her2+, 5 ER + and 4 TNBC samples was analyzed with one way ANOVA to identify a list of significant genes with p-value < 0.05 used as a significance threshold. Expression patterns of the significant genes were used for Principal Component Analysis. Projection of training and testing set samples on the first two principal components was used to visualize relationship between samples.
After the validation, a final list of significant genes differentially expressed between three classes of samples (Her2+, ER + and TNBC) was determined by using one way ANOVA on the full set of samples, except for the one Her2+/ER + sample (TB148). False discovery rate (FDR) was determined according to published protocol . Significance for genes between each pair of groups was determined by Tukey post-hoc test. P-value <0.05 was set as a significance threshold.
Identification of biological functions and pathways overrepresented in any gene list was done using DAVID  and Ingenuity Pathway Analysis (IPA) software (Ingenuity Systems, Redwood City, CA). DAVID results were restricted to gene ontology (GO) terms, KEGG, and BIOCARTA pathways and Swiss-Prot keyword enrichments and filtered to satisfy FDR <5% and fold enrichment >2 criteria. Significance of IPA results was defined by Benjamini-Hochberg corrected for multiple testing p-value < 0.05.
Heatmap was generated for a list of the 44 significant genes (with a fold change > 2) that distinguish Her2+ CAFs from both ER + and TNBC derived CAFs. Genes were hierarchically clustered using Spearman correlation distance and complete linkage. Heatmap color intensities were proportional to a value calculated as a ratio between the gene expression in a single sample and the geometric mean expression of the gene across all samples.
Expression of six genes, ITGA3, ITGA5, OXTR, WNT5B, BCAR1 and FZD1, as well as 3 endogenous controls (ec) RPL19, TBP and UBA5 were assessed by qRT-PCR in triplicates. Median Ct values for each gene were used for ΔΔCt analysis, where ΔCt was calculated against average Ct of the three endogenous controls and ΔΔCt calculated as difference between average ΔCt values of compared groups. Final fold change between a pair of groups was calculated as 2ΔΔCt. Significance of the difference between two groups was tested by two-tail t-test on ΔCt values. For comparison with expression values from microarrays, corrected for loading bias absolute expression values E for each gene G were calculated as follows: E = AE G /(AE ec /avg(AE ec )), where absolute expression AE G = 240-Ct , AE ec is an average AE between three endogenous controls and avg(AE ec ) is an average of AE ec taken across all samples. Expression values were then normalized for microarray and qRT-PCR data separately over three group average absolute expression values.
The migration properties of T47D (ATCC), a breast cancer cell line, known to have low migratory properties , was evaluated in the presence or absence of CAFs derived from ER, TNBC, and Her2+ breast cancer using a transwell assay. CAFs (1×104 cells) from each of the three subtypes were seeded in 100 μl of DMEM containing 1% serum medium in the lower well of a Transwell chamber (Costar, Inc.) with 8 μm pore size polycarbonate filters and left to attach for 90mins. As control, medium containing no CAFs was placed in the lower well. T47D (1×104 cells) were then seeded onto the upper chamber in 1% serum medium. Transwell chambers were incubated for 48 hours at 37°C and 5% CO2. Membranes were stained with DAPI (Invitrogen) for 15 min, rinsed with PBS and fixed with 10% buffered formalin (Fisher Scientific, SF100-20) for 15 min before imaging. The number of T47D cells that migrated onto the underside of the membrane was counted in 5 fields using a Nikon TE2000 inverted microscope at 10× magnification and plotted. Statistical evaluation was performed using Graph Pad Prism (GraphPad Software, Inc.).
The clinical characteristics of the study cohort are summarized in Table 2. Detailed clinical characteristics of each tumor are provided in Additional file 1: Table S1. No significant differences were noted among the three subgroups, except for tumor grade (Table 2). The morphology of CAFs isolated from the 3 different breast cancer subtypes was similar (Figure 1). Further phenotypic characterization using flow cytometry analysis demonstrated that >95% of these cells expressed fibroblast activation protein (FAP), a previously identified marker of cancer associated fibroblasts [25–28]. Moreover, >99% of the cells were negative for the epithelial cell adhesion molecule (EpCAM), a breast cancer epithelial cell surface marker ; CD31, also known as platelet endothelial cell adhesion molecule (PECAM-1), an endothelial cell marker, and CD45, a pan-leukocyte marker (Figure 2, lower panel). Moreover, these CAFs uniformly expressed vimentin and collagen by immunohistochemistry (data not shown).
Relationship between Her2+, ER + and TNBC classes of samples visualized by Principal Component Analysis (PCA) on the training set samples using expression of genes differentially expressed between the three classes. A. Training set samples B. Projection of testing set samples on the first and second principal components derived from the training set. White square in dark grey diamond indicates tested sample with double diagnosis Her2+/ER +.
RNA isolated from the early passage CAFs were assayed for gene expression and randomly assigned to two sample sets, namely, training and testing sets (Table 1) to perform independent validation. Using one-way ANOVA on the training set (4 TNBC samples, 5 ER + samples and 3 Her2+ samples)), we identified 782 genes that were differentially expressed between TNBC, ER + and Her2+ samples (p-value < 0.05). In order to visualize the relationships between the sample types, we performed unsupervised Principal Component Analysis using the 782 significant genes (Figure 2A). This type of plot reflects the similarities and differences between all samples in relation to the 782 significant genes. It should be noted that the first principal component plotted on the X axis accounts for 49% of the variation in the data and indicates that there are significant differences between the CAFs derived from the Her2+ cancers and both the TNBC and ER + breast cancers, as these samples are equally separated from the Her2+ samples along the X axis. The second principal component plotted on the Y axis accounts for only 14% of the gene expression variation between all samples. It captures putative differences between the ER + and TNBC samples and indicates that the expression profiles are much more similar between these two subtypes.
Venn diagram for genes common between three pair-wise comparisons of Her2+, ER + and TNBC classes of samples.
We then determined whether the training set principal components could also distinguish the new Her2+, ER + and TNBC patient samples thus validating our initial observations. Figure 2A shows the separation of the 12 samples representing the 3 original sample types in the training set that we used to select the significant genes that defined this separation. Figure 2B confirms these genes also identify the subtype differences in new samples analyzed as an independent validation set and included two new Her2+ samples and t two new ER + and two new TNBC samples. The new Her2+ samples clearly cluster with the Her2+ samples in the training set while the new ER + and TNBC samples once again cluster with the ER + and TNBC training set samples. Although the ER + and TNBC derived CAFs appear to self segregate along the 2nd principal component in the training set (Figure 2A), no significant differences in gene expression were detected between the ER + and TNBC CAFs in the testing set (Figure 2B). This indicates that there is a high degree of gene expression similarity in the CAFs associated with the ER + and TNBC cancer subtypes.
It should also be noted that new sample TB148, which is both Her2+ and ER+, co-segregates with the Her2+ samples which were all ER- (Figure 2B), indicating the presence of a gene expression profile more similar to the Her2+ CAFs and not the ER + CAF sample group. This indicates a dominance of Her2+ CAF gene expression signature over ER + CAF signature.
We also combined the expression data for all samples (except for the Her2+/ER + TB148) to take advantage of the larger sample size and ran one way ANOVA to define a final list of significant genes differentially expressed between Her2+, ER + and TNBC in the larger data set. We found 1829 differentially expressed genes with p-value < 0.05 and estimated false discovery rate of 28%. When the relationships between the different CAF subtypes were reassessed using Principal Component Analysis with the new gene set, we found the same cancer subtype specific differences as demonstrated on training subset (Figure 2A).
The number of significant genes identified by pair-wise comparisons (Tukey post-hoc test) between the three classes of patient samples, i.e. Her2+ vs ER+, Her2+ vs. TNBC and ER + vs TNBC samples, are presented in the Venn diagram in Figure 3. These results quantify the visual interpretation of Principal Component Analysis demonstrating that while 1,800 genes were significantly differentially expressed between Her2+ and either ER + or TNBC, only 118 genes were significantly different between ER + and TNBC derived CAFs. Further studies with increased number of samples for ER + and TNBC derived CAFs will be required to identify genes that can discrimi-nate those 2 classes, if they exist. A gene expression heat map for the 44 most changed unique genes (fold change > 2) which were common to the Her2+ vs ER + and Her2 + vs TNBC comparisons are shown in Figure 4.
Heat map of expression for 44 genes with the greatest differences between Her2+ vs. ER + and Her2+ vs. TNBC comparisons. FC = fold change from geometrical mean of expression across all samples.
qRT-PCR validation. qPCR was used to validate microarray results for 6 genes found to be significantly different in either Her2+ vs ER+, Her2+ vs TNBC or ER + vs TNBC comparison in microarrays data. Expression for arrays and qPCR were normalized separately over average value across absolute expression for Her2 +, ER + and TNBC groups. Error bars represent standard error of mean for the group.
We compared the two significant gene lists for Her2+ vs ER + and Her2+ vs TNBC to identify functions or pathways that might be over-represented among the differentially expressed genes. Results with DAVID software analyses  are shown in Additional file 2: Table S2 for the Her2+ vs ER + 1253 significant genes, and in Additional file 3: Table S3 for Her2+ vs TNBC 1035 significant genes. Enrichment of nine functional categories associated with cytoskeleton and extracellular matrix were found to be significant in both comparisons.
Ingenuity pathway analysis was done for a list of 615 genes common between Her2+ vs ER + and Her2+ vs. TNBC comparisons. A list of significantly enriched canonical pathways is presented in Table 3. Pathways involving extracellular matrix/integrin signaling were found to be significantly up-regulated in CAFs derived from Her2+ cancer, further supporting the DAVID results. It should be noted that 92% (61 of the 66 unique) of the genes associated with the ingenuity pathways are upregulated in Her2+ supporting the hypothesis that those pathways are more active in CAFs derived from Her2+ breast cancer as compared to those derived from the ER + and TNBC breast cancers.
pval = Benjamini-Hochberg corrected p-value, P = total number of genes known to be involved in the pathway, L = number of genes from the pathway that were also in the list of significant genes. ↑ = number of genes significantly upregulated in Her2+, ↓ = number of genes significantly downregulated in Her2+. The 18 significantly enriched pathways share 66 unique genes with 61 of those upregulated in Her2+ compared to ER + and TNBC.
To confirm differential gene expression levels in the three breast cancer subtypes, Her2+, ER + and TNBC, we selected 6 genes (ITGA3, ITGA5, OXTR, WNT5B, BCAR1, FZD1) with significantly different levels of expression based on our microarray studies and validated their expression levels by qRT-PCR. Fold changes in expression based on the arrays ranged from 1.5 fold to 6.9 fold. Five of the 6 genes that were found to be expressed at higher levels in the Her2+ samples were also significantly different in the Her2+/ER + qRT-PCR comparison; and 4 of those 5 genes that were significantly different in the Her2+/TNBC array comparison were also significantly different by qRT-PCR comparison (Figure 5 and Additional file 4: Table S4). Expression ratios by qRT-PCR were highly consistent with array values and overall somewhat higher by qRT-PCR as expected. One gene, FZD1, which was expressed at lower levels in CAFs derived from Her2+ breast cancer by array analyses, was also significantly lower by qRT-PCR in the Her2/TNBC comparison but was not significantly different in the ER/TNBC comparison (P = 0.2) although fold change values were similar by qRT-PCR (TNBC/ER + = 1.33 for microarrays and 1.39 for qPCR).
CAFs derived from Her2+ breast cancer significantly enhances the migration of T47D cells in vitro. In vitro transwell assays comparing T47D migration in the absence (orange) or presence of CAFs isolated from ER (black), Her2 (blue) and TNBC (green) primary human breast cancer tumors were performed. Each experiment was performed in duplicates using CAFs derived from at least two different patients. One CAF cell line of each subtype was tested in 2 independent experiments (open vs. closed circles). The second CAF cell line of each subtype (squares) was tested in duplicate in one independent experiment, for a total of 6 tests. Lines show mean ± SEM.
To explore whether CAFs derived from various breast cancer subtypes can differentially enhance the migratory phenotype of breast cancer cells, we performed in vitro transwell assays comparing the migration of breast cancer cells cultured in the presence or absence of CAFs isolated from ER+, Her2+ and TNBC. The number of migrated T47 cells onto the membrane surface that was facing the lower chamber was counted. Results were analyzed by unpaired Kruskal-Wallis test. The level of statistical significance was taken as P < 0.05. As our gene expression profile results have predicted, CAFs derived from Her2+ breast cancer significantly enhanced the migration of T47D (Figure 6).
Robust evidence is now available that underscores the role of CAFs in tumor progression [8, 28–33]. Previous gene expression profile analyses comparing CAFs and fibroblasts derived from matched normal adjacent breast tissues have demonstrated significant differences between the CAF and their normal counterparts but, to the best of our knowledge, no prior studies have addressed whether CAFs derived from various breast cancer subtypes harbor subtype specific gene expression signatures. In this study we demonstrate for the first time that CAFs from several breast cancer subtypes exhibit subtype-specific gene expression profiles. Specifically, we show that the gene expression profile of CAFs derived from Her2+ breast cancers are significantly different from CAFs derived from ER + or TNBC breast cancers.
Heterogeneity among fibroblasts has been described in various organ sites including lung, skin, sclera and orbit . Furthermore, Sugimoto and coworkers demonstrated that the expression of various fibroblast markers are heterogeneous within the tumor stroma in mouse breast and pancreatic tumor models using immunohistochemical analyses . Several studies have generated gene expression profiles from breast cancer-associated fibroblasts but none of these studies have stratified their results based on tumor subtypes. Work by Allinen and coworkers evaluated gene expression profiles of breast cancer stromal cells which were isolated by negatively selecting out epithelial cells, lymphocytes and endothelial cells . Work described by Singer et al. compared gene expression profiles of stromal fibroblasts derived from 10 invasive breast cancers with stromal fibroblasts derived from normal breast tissues of 10 women undergoing breast reduction surgery . Their results demonstrated increased expression of tumor promotion-associated genes in the pooled CAFs. Work by Bauer et al. (2010) evaluated gene expression profiles of fibroblasts derived from 6 matched breast cancers and adjacent normal breast tissues  and found distinct differences between CAFs and normal fibroblasts, specifically in genes related to paracrine or intracellular signaling, transcriptional regulation, extracellular matrix and cell adhesion/migration. However, all of the above studies were not designed to test subtype specific differences in CAFs due to these studies' relatively small sample size. In addition, when tumor subtype data were reported, the less common breast cancer subtypes, i.e., Her2+ or TNBC cancer, were underrepresented.
Our results showed that CAFs derived from Her2+ breast cancers significantly up-regulated pathways associated with actin cytoskeleton and integrin signaling (Table 3). Integrins mediate cell attachment with extracellular matrix (ECM) to provide traction necessary for cell motility and invasion. These upregulated signaling pathways may have contributed to the elevated migratory phenotype of breast cancer cells (T47D) in our in vitro transwell assays (Figure 6).
The extracellular matrix and integrins collaborate to regulate gene expression associated with cell growth, differentiation and survival; all of which are deregulated during cancer progression and metastasis. A recent study using a three-dimensional squamous cell carcinoma (SCC)/fibroblast co-culture model elegantly demonstrated the role of three genes, integrin α3, integrin α5 and Rho, in promoting a fibroblast-led collective invasion of SCC cells into the extracellular matrix . Interestingly, all three genes were significantly up-regulated in CAFs derived from Her2+ breast cancer with integrin signaling as the second most enriched pathway (Table 3). Moreover, many of the genes and pathways downstream of integrin signaling are also significantly upregulated in Her2+ CAFs. These include focal adhesion kinase (FAK), Rac and Rho signaling pathways as well as several members of the mitogen-activated protein kinases (MAPKs), further underscoring the importance of integrin signaling in CAF. In addition to the well-established role of integrins in migration and invasion, integrins can also regulate cell proliferation, including mammary gland proliferation  through integrin-linked kinase (ILK) , which was also noted to be significantly upregulated in HER2+ derived CAFs. These characteristic differences in CAFs derived from Her2+ breast cancer may contribute to the aggressiveness of this particular breast cancer subtype which is known to have an increased propensity for local and distant recurrence . In addition, the sites of distant metastasis appear to differ according to breast cancer subtype with Her2+ breast cancer having a higher rate of brain, liver, and lung metastases than ER + breast cancer . The role of CAF in contributing to a subtype-specific trophism for the various distant metastatic sites is unknown.
Gene expression profile differences between CAFs derived from ER + and TNBC breast cancer were less pronounced and we were unable to confirm them with independent validation set using the limited sample numbers (Figure 2B). While it is possible that true differences may exist among these two subtypes, a larger number of samples would be required to find those differences with an acceptable false discovery rate.
Our results show that subtype specific changes exist in CAFs derived from breast cancer. In the case of Her2+ breast cancer, a more aggressive breast cancer subtype with known increased risk of local and distant recurrence, CAFs may augment the invasive properties of the tumor cells via pathways associated with cytoskeleton and integrin signaling. Our findings also provided molecular evidence supporting a recently proposed tumor-stroma co-evolution hypothesis which suggested that the tumor microenvironment, e.g. CAFs, may adopt specific changes to optimize the survival/propagation of a specific tumor cell type . Whether these programmatic differences in CAFs result from epigenetic changes or whether these differences are due to heterogeneity within the CAF population, i.e. proportion of resident fibroblasts vs. recruited fibroblasts, or fibroblasts derived from epithelial mesenchymal transition are unknown. In addition, whether CAFs contribute to tumor progression in a subtype specific manner is unknown. How CAFs and other components of the tumor microenvironment drive or are being driven by the tumor cells to promote the propagation and maintenance of a specific tumor subtype will be the subject of future work.
Louise C Showe and Ellen Puré contributed equally to this work.
The authors thank the Tumor Tissue and Biospecimen Bank (TTAB) of the Abramson Cancer Center, Perelman School of Medicine of University of Pennsylvania, for assisting in tumor tissue collection.
This research was, in part, funded by the NCI Cancer Center Support Grant (2-P30-CA-016520-35) (J. Tchou), and the Linda and Paul Richardson Breast Cancer Research Funds (J. Tchou).
JT, AVK and LS designed the study; JT, AVK, LC, CS performed the experiments described in this study; JT, AVK, LC, MH, LS and EP contributed to the writing of the manuscript. All authors read and approved the final manuscript.


Do you love roses? Luscious rose gardens add a beautiful atmosphere to any home and garden. Even beginners can grow roses with little trouble when learn the basics of maintenance. There are some gorgeous ideas for backyard rose gardening. Although there are many steps to growing a beautiful rose garden, even a beginner can do it. Here you will find some rose gardening tips for the beginner.
– First, choose your spot. Roses love sunlight; therefore, they grow best when getting at least 8 hours of sunshine daily. A full day of sunlight is best; however, morning sun is preferable to afternoon sun.
– Prepare the soil for your roses well. Although, roses are not fragile flowers, they thrive in good draining soil, with a pH level about 6 or 7. Roses flourish in soils rich in organic materials.
– Keep in mind that soil that does not drain well, leave plants open to the vulnerability of disease and roots rot.
– Add organic materials such as compost, peat moss and manure.
– Fertilizers come in various choices between organic and chemical fertilizers.
– Allow the soil to rest a few days as it settles.
There are various choices available when buying roses, such as colors and the different breeds as well as the types such as bushes, climbing roses, grafted and root roses. However, you should be aware that the difference between grafted and root roses, is that grafted roses are those which are weaker roses that have been grafted onto a much hardier rootstock rose. Grafted roses do have the disadvantage of coming back as a different type of rose once it dies back in the winter. It is then the rootstock rose, which is generally not appealing. However, there are rose varieties that are available as grafted roses, such as tea roses.
The last step in rose gardening is to plant the roses. It is essential to take care not to cause any damage to the roses, since any cuts cause the plant to become even more susceptible to disease, which makes it harder to care for. There are two types of roses available for purchase, they are container and bare root roses. The roots of bare root roses need to be moist. You can place the roots in a bucket with a solution of water and organic materials, such as fish emulsion, to reach the desired moistness. Simply pull out the roots and then cut off any soft, mushy areas.
First, you must dig holes where you will plant the roses, for either container or bare root roses. In the bottom of each of the holes, you should apply some potassium, either a banana peel or an additive from your local gardening center. Locate the root bulb; this area joins the bulb to the root of the plant stem. In milder climates, do not bury this; however, in cooler climates it is best to place them at least 3 inches underground.
Newly planted rose plants need a bit of pampering. This means you should water them daily for 3 weeks, as long as the soil has good drainage. Doing so will encourage your roses to flourish. However, you should keep in mind while watering your roses, to water the soil, while avoiding getting the leaves and flowers wet. Moist foliage encourages the growth of fungi. Many roses need pruning after they have grown, which in turn encourages new growth.
When you love roses and are searching for great gardening tips, you should visit your neighbors, the local gardening center or the internet, as these are all great areas to learn. With a bit of effort and patience, your rose garden will become an amazing place of beauty.
Check out The Joy of Rose Gardening now at http://www.roseflowergarden.com, to find great articles, advice, tips and much more on rose gardening topics.


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prandisound » Pop » Leona - Leona
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FLAC size: 1397 mb | MP3 size: 1442 mb | WMA size: 1253 mb
Leona Naess is a 2003 self-titled album by Leona Naess. Leona Naess is Naess' third album, and the first of hers to be produced Geffen Records. One of the tracks, Christmas, also appeared on the Music from the OC: Mix 3, and the album reached 43 on the TopHeatseekers chart in 2003.
Leona Berlin (2018) - download the album and listen online. On this page you can listen to the album, get information about the album, see the list of songs and much more. Listen online and stay in a good mood. Org Album: Leona Berlin (2018).
Spirit is the debut studio album by British singer-songwriter Leona Lewis, released by Syco Music in November 2007 in the United Kingdom and Ireland, followed by a worldwide release during early 2008. After winning the third series of reality singing contest The X Factor in December 2006, Lewis began recording the album in Sweden, the United Kingdom and United States, working with a variety of writers and producers
I Am is the fifth studio album by British singer and songwriter Leona Lewis. It was released on 11 September 2015, by Island Records after Lewis took leave from Syco Music in June 2014. Recording began in mid 2013, during the production of Lewis's first Christmas and fourth studio album, Christmas, with Love (2013). Recording sessions resumed in January 2014 with producer Poet, Toby Gad and Wayne Wilkins
The problem for Lewis on her follow-up album is that while this is the exact point at which she should be marking out her own territory, Echo feels and sounds even more a part of the X Factory than her debut album. There's still nothing not to love and, talentwise, the girl from Hackney is up there with anyone you care to name from Hollywood. But Echo plays it so safe it could have been made by any female singer of the past 25 years. As air brushed as its cover art, this is Leona as generic diva when she could be so much more. Leona Lewis Music Sony X Factor.
Leona (レオナ, Reona), also known under her codename Leona Heidern (レオナ・ハイデルン, Reona Haiderun), is a character introduced in The King of Fighters '96 as the new member to the Ikari Team. Her official nickname is The Silent Soldier (サイレント ソルジャー). Aside from the main body of the King of Fighters series, Leona is also featured in a one-shot chapter in the spin-off manga, The King of Fighters: Kyo and has her own character image album.
1 Ukolébavka 4:12
2 Děj Se Děj 4:07
3 Pár Tvejch Očí 4:01
4 Beránek 4:23
5 Píseň Ke Kuchyňskému Stolu 3:37
6 Průhledná 4:35
7 Imrvére 4:01
8 Klíč 4:33
9 Schody Z Písku 4:05
10 Dej Na Mě 3:54
11 Prošla Tmou 4:15
MM 0612-2 Leona* Leona (CD, Album, Enh, RE, RM) Master Music MM 0612-2 Czech Republic 2006
Related to Leona - Leona
Various - Als Jij Voelt Wat Ik Voel download mp3
Leona Lewis - Happy Remixes download mp3
Leona Lewis - Bleeding Love download mp3
Leona - Zomer Op Je Radio / Summer On Your Radio download mp3
Leona Lewis - The Labyrinth Tour (Live From The O2) download mp3
Lee Andrews & The Hearts - Lonely Room / Leona download mp3
Kontraband Milana Svobody, Various - Vánoční Písně & Koledy download mp3
Michael Bolton Featuring Leona Lewis - Ain't No Mountain High Enough download mp3
Leona Lewis - Christmas, With Love download mp3
Leona Lewis - Forgive Me download mp3
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redpajama_set_name: RedPajamaCommonCrawl


text:
Wynnum residents refuse to live with toxic coal dust
Concerned Wynnum residents have called on the State Government to demand the coal industry and Queensland Rail cover coal stockpiles and train wagons passing through densely populated areas of Queensland.
Clean Air Queensland (CAQ hosted a community barbeque yesterday at Wynnum to highlight the issue of toxic coal dust in local homes, schools and workplaces.
Clean Air Queensland (CAQ) spokesperson, Michael Kane told the gathering that the government, mining companies and Queensland Rail were ignoring their responsibility not to endanger the health of Queenslanders.
"The people of Wynnum and others living along coal train lines from Brisbane to the port have had enough and are demanding action from this government to stand up for them against the powerful coal industry," said Mr Kane.
Wynnum resident of 43 years, Colleen Durnian spoke of concerns for her husband's health as his asthma was worsening as the coal dust in and around their house increased.
"The front of my house is black and it's a struggle to keep the coal dust out," Mrs Durnian said. "His asthma is increasingly getting worse and we can't understand why they just won't cover the coal to stop the problem."
Exposure to particulate pollution from substances like coal dust is known to cause asthma, lung cancer and respiratory illnesses.
Public Health researcher, Associate Professor Adrian Barnett of the Queenslander University of Technology backed the results of a CAQ study - ''Health Hazard in our Suburbs' – that showed coal trains were emitting alarming amounts of pollution as they passed close to homes, schools and workplaces.
Mr Kane has sent the report, released last week, to Premier Annastacia Palaszczuk, Environment Minister Miles, Natural Resources and Mines Minister Lynham and the Speaker, Peter Wellington.
The CAQ study that found that coal trains passing through Wynnum emitted up to 900% higher pollution readings than normal levels. Towns including Toowoomba and Ipswich and 21 Brisbane suburbs were impacted.


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Virus Removal in Northern Kentucky. It's a jungle out there - with everything from viruses, to hackers, to spyware and malware infections.
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Community Computer Services has been in business for 25 years. We are located in Alexandria Kentucky. We specialize in computer repair, sales, upgrades, networking, laptop repair, and mac repair. If your computer is running slow or having pop-ups then choose us, an honest, family-owned local small business. We work on Dell, HP, Acer, Toshiba, IBM, and much more. We service many areas such as Alexandria, Cold Spring, Highland Heights, Wilder, Southgate, FT. Thomas, Falmouth, all of Boone, Kenton, and Campbell County.
We charge $45.00 to diagnose your computer. If there is work to be done then we apply that towards the repair.
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I had a really exceptional experience with the staff and would highly recommended to my peers and colleagues!
The free repair guide for everything Mac, and Windows related.


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The Illegal IRS Rule to Increase Taxes & Spending under ObamaCare: Our Response to Timothy Jost
By Michael F. Cannon
Jonathan Adler and I have a post at the at the Health Affairs blog where we respond to Timothy Jost's critique of our working paper, "Taxation without Representation: the Illegal IRS Rule to Expand Tax Credits under the PPACA." Jost has been our most tenacious (if not most consistent) critic.
Here's an excerpt. Keep in mind that although we say "tax credits," government spending accounts for about 80 percent of the money involved. Which is a lot: the cost of this illegal IRS rule could be in the hundreds of billions of dollars.
The dispute is over whether the [Patient Protection and Affordable Care] Act authorizes the IRS to provide tax credits only in Exchanges established by states (under Section 1311) or also in Exchanges established by the federal government (under Section 1321). Three facts are key to this dispute.
First, both sides acknowledge that the statutory language governing eligibility for tax credits is clear and unambiguous. The Act provides that taxpayers are eligible for tax credits if they purchase a health plan through "an Exchange established by the State under section 1311." That language clearly authorizes tax credits only in state‐established Exchanges, and the Act employs or refers to that language no less than six times when authorizing tax credits. There is no parallel language anywhere in the statute authorizing the IRS to offer tax credits through federal Exchanges established under Section 1321.
Second, there is nothing in the statute that conflicts with the plain meaning of that language. Indeed, the rest of the statute supports that plain meaning. Nor has anyone identified anything in the law's legislative history that conflicts with that language. The only statement anyone has found on this point shows the statutory language was intentional. During congressional debate, the bill's lead author, Senate Finance Committee chairman Max Baucus (D‑MT), explained that the bill conditions tax credits on the establishment of a state‐run Exchange.
Third, even though some members of Congress and the President might have preferred a law that authorized tax credits in federal Exchanges, they nevertheless enacted a law that did not. Many advocates of health care reform urged passage of the Senate bill even though there were parts of the bill they did not like, and knowing full well that not all defects could or would be fixed through the reconciliation process. Congress amended the sections of the Senate bill that authorize tax credits and cost‐sharing subsidies a total of 12 times through the reconciliation process, but left the language limiting tax credits to state‐established Exchanges undisturbed. Again, many of those amendments support the clear meaning of that language, and none of them conflict with it.
And yet, in late May the IRS finalized a rule that will issue tax credits — and therefore will trigger cost‐sharing subsidies and employer‐mandate penalties — through federal Exchanges, contrary to the plain language of the statute. It is our contention that this rule is illegal.
We invite everyone to read our working paper alongside Jost's post, and our reply, and to decide for themselves whether the IRS is breaking the law.
You can also watch Jost and me testify before Congress on the IRS rule tomorrow at 9am ET in room 2154 of the Rayburn House Office Building.
General, Government and Politics, Health Care, Constitution, the Law, and the Courts


Anyone that works within any of the following fields is entitled to 10% off their complete glasses price as long as they have spent £49 or more on frames and also full priced lenses.
2. Emergency Services (Police, Fire or Ambulance).
3. Member of the UK Armed Forces.
Proof of employment in these fields is required to receive entitlement.


Harold & Kumar alum Kal Penn is set to co-star opposite Tyler Labine in the upcoming third season of Hulu's original comedy series Deadbeat.
The supernatural comedy centers on Kevin Pacalioglu (Labine), a hapless but gifted medium who will go to any lengths to help New York's ghosts settle their unfinished business. Penn will play Clyde, stoner entrepreneur extraordinaire and all-around good dude, who becomes Pac's business partner.
Deadbeat starts production on its third season in New York on Monday. The third season is slated to premiere on Hulu in spring 2016.
Industry Entertainment-repped Penn will next be seen in feature Stereotypically You.
Created by Cody Heller and Brett Konner (Wilfred, The Inbetweeners), Deadbeat is produced by Lionsgate Television in association with Brad Pitt's Plan B Entertainment and Dakota Pictures.


The Nevada Supreme Court published a landmark ruling dealing with premises liability on December 27, 2012. The Court, in Foster v. Costco, 128 Nev. Adv. Op. 71, took a long look at the history of landowner liability in Nevada and held that the mere fact that a condition may be open and obvious does not automatically negate a landowner's duty of care to someone injured on his property by the allegedly open and obvious condition.
Defendants in Nevada premises cases have often moved for summary judgment (arguing that no genuine issues of material fact exist and so the case can be decided as a matter of law) in "open and obvious" cases. This will likely stop nearly completely with the Court's holding in Foster.
other risks to entrants on the land when any of the affirmative duties is . . . applicable.
This duty is extended to all entrants on land, not just those invited. Landowners bear a general duty of reasonable care to all entrants, stated the Court, regardless of the open and obvious nature of dangerous conditions.
The duty must be analyzed with regard to foreseeability and gravity of harm, and the feasibility and availability of alternative conduct that would have prevented the harm.
In considering whether reasonable care was taken, the fact-finder must also take into account the surrounding circumstances, such as whether the landowner had reason to suspect that the entrant would proceed despite a known or obvious danger.
Separate from but related to the reasonable care assessment is consideration of the entrant's actions and whether he or she failed to exercise reasonable self-protection in encountering the danger.
The Court determined in this case that Costco is not free from liability under Nevada law solely because the danger of the pallet in its aisle may have been open and obvious to Foster. A jury could reasonably believe that Foster walked down the paper goods aisle without observing the dangerous condition (a pallet extending into the aisle). Even if a jury found the risk to be open and obvious, it must also decide whether Costco breached its duty to allow the condition to exist. And a jury must also be allowed to examine Foster's own negligence, if any. The Court therefore remanded back to the District Court for further review.


The LTMP 2040 is a long-term land transport master plan to shape the future of land transport to 2040, and possibly beyond.
In LTMP 2008 and 2013, we set out our vision for a people-centred land transport system. We promised more connections, better service and a transport system that supports a liveable and inclusive community.
A convenient, connected, and fast land transport system.
More inclusive infrastructure like ramps and lifts, and a sense of shared and individual responsibility in the community.
Healthier lifestyles, more appealing walking and cycling options, as well as cleaner and quieter roads.
The public engagement report can be found here.
We are deeply grateful to the many Singaporeans who took the time to share their views, comments, and suggestions with us.
Enabling Singaporeans to Enjoy 20-minute towns and a 45-minute city on public, active and shared modes of transport.
The LTMP Advisory Panel's recommendations report can be found here. The Government has accepted these recommendations and will release a fuller set of measures in due course.


Landon Robinson joined FMK this week. Welcome to the team!
We're thrilled to add yet another member to the FMK team this week!
Please join us in welcoming, Akielah Wiggins.
FMK is expanding! We're glad to welcome Amelia Wilson to the team.


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Home Software Should I Get a PS4 or Xbox One?
Should I Get a PS4 or Xbox One?
By Luis LeonDec 07, 2015, 06:31 amComments Off on Should I Get a PS4 or Xbox One?
With both of the leading consoles vying for supremacy over nearly two full years, each has had a chance to iron out a few preliminary wrinkles in order to deliver the best all-around gaming experience. Each has a distinct approach, and the characteristic differences between each console can make one console a preferable choice for a given user.
Sony's PlayStation 4 is a much more streamlined console than the Xbox One, taking the form of a slim rectangle as opposed to the much larger Xbox One, which weighs in at over 13 pounds and features a larger, less portable power cord as well. However, this larger chassis is intended to address a common concern with the previous generation Xbox 360, which regularly experienced overheating.
The PS4 is marketed chiefly at its established market of dedicated gamers, equipped with a more powerful graphics processing unit that allows for more cinematic gameplay. Another feature, PlayStation TV, allows users to stream and play games over a Wi-Fi connection. The PS4 also allows gamers to share games or even play cooperatively over a remote connection with SharePlay, and gamers can buy PlayStation 4 games from Harvey Norman online for immediate delivery.
On the other hand, one of the most versatile features of the well-rounded Xbox is its bundled motion detecting camera, the Kinect. Unlike the PlayStation Camera of the PS4, the Kinect is fully integrated into the user experience of the Xbox One, enabling input in system setting as well as control during gameplay. The Xbox One also has an HDMI input and native 3D Blu-Ray support as standard features to further distinguish it from the base model PS4.
The PlayStation4 employs a minimalistic user interface designed with maximum ease of use for new users in mind. It also releases periodic software update with its PlayStation Plus service, although the updates tend to be more gaming focused than directed at expanding the PS4's capabilities as a home entertainment system.
In contrast, Microsoft designed the interface of the Xbox One to be similar to its modern Windows platforms such as Windows 10, delivering a desktop style experience. A subscription to Xbox Live gives users access to monthly updates as long as they remain an active subscriber, and past updates have delivered capabilities including smart TV functionality and advanced storage features, making it an excellent choice for the experienced gamer.
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Artists Control Each Other's Body Parts with Their Minds
Artist Diva Helmy and neuroscientist Greg Gage make unbelievable art out of a DIY neuroscience interface kit.
by DJ Pangburn
Images courtesy the artist
In visual effects master Douglas Trumbull's (2001: A Space Odyssey) last feature film, the 1983 proto-cyberpunk Brainstorm, characters create an interface that allows humans to feel the sensations of others. Norway-based artist Diva Helmy's use of the Human to Human Interface (HHsI) isn't nearly as precise as this fictional one, but HHsI does allow humans to manipulate and control others, to a certain degree.
With technical development and production by Greg Gage, HHsI allows one individual to wirelessly control the arms of multiple bodies with their brain signals. In a recently released short film titled The Controller, Helmy shows one person controlling another, then another person controlling five. While electrodes are easily seen in the video, HHsI also features an Arduino board, MuscleSpiker shield (which allows users to control things with their muscles), a TENS unit (nerve stimulator), 9-volt batteries, and radio frequency modules.
The project was begun when Helmy and Gage spoke over Skype last summer about a possible collaboration. The Human to Human Interface already existed, but the two wanted to modify it so it could be used at a larger scale in an artistic context. It now features one controller and ten controlled user units.
"The idea was to create an experience in which gallery visitors could use the device and form a physical and wireless network of electrical discharge between each other, one that forced the controlled users to simultaneously mirror the behavior of the controller," Helmy tells The Creators Project. "The interface presents an exploration into the world of neuroprosthetics and forced control of others' nervous systems through technology."
Helmy and Gage are interested in the notion of losing control over one's own body, and what happens as a result. The Controller, like a science experiment of sorts, is collaborative: two people perform one task—such as playing music—through one body.
"The device transfers the brain signal of one person to the ulnar nerves in the arms of several individuals," Helmy explains. "The first human's EMG signal (neurological activity in muscle cells) is recorded as they move their arm. The value of the signal activates the nerve stimulators via radio frequency which then sends an electrical sensation through the arms of the controlled bodies forcing them to move based on when the first individual sends a brain signal to move their own arm."
All arms move simultaneously based on controller's brain signal. The wireless signals can be sent between the units up to a distance of 65'. And Helmy says that the technology could support 100 controlled users.
As neat as it is, there is something about the device that might stoke the fears of the paranoid. Keep in mind, though, that this neat technological voodoo kit requires several components. So it's not as if unseen forces will be controlling us anytime soon, if ever. Well, at least not physically.
Click here to see more of Diva Helmy's work, and here to check out more of Greg Gage's electrophysiology work.
Change Your Sex in Virtual Reality
Amputee Gets Two Mind-Controlled Robotic Arms
Artist Manipulates Water With The Power Of Her Mind
Greg Gage
DIY neuroscience interface
Diva Helmy
HHsI
Human to Human Interface


Balls are essential and strategic elements in the production of industrial components and the smooth functioning of numerous products, vehicles and consumables.
CIMAP has acquired experience and a reputation in supplying balls for all these applications since its foundation in 1958.
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Article Summary: "This article is all about how a person can be motivated by his or her company in many ways. The article is a part of my research conducted to give a better idea of a strong reward system. I am writing this article after a long gap to understand how you viewers take such a topic in a negative or positive way. This is just a small.."
We all know that an organization is a well planned management to conduct business internally as well as externally. According to my research an organization can only grow when they have a well planned reward system to motivate employees. It is important that a company should recognize the efforts put by its employees. Many believe that using a positive approach for encouragement can produce better work and quality, but this is only possible when an employee is rewarded for his efforts. Every company should get to know their employees needs and desires. A well planned reward system needs to be introduce. This will motivate employees to do better. A company should introduce rewards such as hike in salary, promotions, parties or a praise to recognize a persons effort would make him or her work in a better flow and quality. Workers that are unhappy intent to work less, produce less quality work, laziness intent to crack through they bones and extra motivation can cause an over confident attitude towards work and fellow employees. You will also see that rewards varies from person to person depending on their needs. You might notice when you get motivated by someone for good work or a deed, you feel proud and like to do the same thing in a proper way with more effort. Incentives is another cycle which can be added to the reward system. A reward can only be earned when an employee helps a company to achieve something. Talking about motivation, challenging tasks should be given where a person a use his or her skills to complete it with perfection and for every challenging task a reward should be given. Not every one can get rewarded, an employee should be given the liberty to choose his or her field of job or task given in which they are good and can be an important asset to the company. Encouraging employees, talking to them about the issue they are facing at workplace, getting an instant solution and make them feel comfortable, motivated and go for their goals can benefit them as well as company. Sometimes an employee may not be happy in what he is doing but he is working just to earn a living. Most important part is security and stability. Every employee looks at a stable job and security. Sometimes these things can also be the cause that a person is not ready to take a step in which they will lose on something. A company should take an important step towards this and bring in that security feeling in a person. If a person knows his job is secured he will intent to work better. An employee should be rewarded with responsibilities and a higher rank or position where he or she can showcase his or her talent. When a company decides to link performance by motivating and encouraging with rewards, this can lead to a strong staff development. The theory where rewards can play an important role in motivating an employee can help in the growth of the company performance. It is a competitive market, and for a company to enter in to business this is an important step to adopt and secure a place of achievement. An effective reward system can attract attention towards a strong build up of an empire. We all look forward to our companies to recognize our skills, talents and to give us an opportunity ahead to achieve our dreams. Understanding of how much each employee puts in all efforts to be wanted as an important asset of the company we all look for a strong reward system and managers to motivate us to do better.
• A Middle-aged Man's Monologue.
• Why Companies Internationalize Their Products?


RRAMAC Connected Systems will exhibit and speak at a number of industry-leading trade shows across the United States. As a provider of remote monitoring solutions and IIoT solutions, Tom Craven, Vice President of Product Strategy for RRAMAC, has been asked to speak at AI Manufacturing. RRAMAC will also exhibit at the AHTD Fall Meeting, ATX Minneapolis, and upcoming Power/mation Technology Fairs.
For free expo passes, conference track discounts, and to learn more about the upcoming presentation and exhibiting engagements, read on.
RRAMAC will be exhibiting at booth #619 at ATX Minneapolis on October 31 – November 1 at the Minneapolis Convention Center. ATX Minneapolis is the largest advanced design and manufacturing event in the Midwest and is right in RRAMAC's backyard with our headquarters located a short distance from the event.
RRAMAC invites you to attend and use our promo code Special619 for FREE Expo Admission and 20% off a Conference Pass. To redeem these offers, register today. Stop by the RRAMAC booth to learn more about our IIoT and remote monitoring solutions and enter to win an echo spot.
Craven will also participate in the ATX Minneapolis matchmaking event to meet potential business partners and connections. If you're interested in learning more about the EdgeScout by RRAMAC solution, please register for a time to connect with us at the matchmaking event. Registration is now open!
Tom Craven will present on "Leveraging the IIoT for Continuous Improvement," on Thursday, September 13 at 2:10pm. Although OEE is one of the gold standards for continuous improvement, many companies are reluctant to implement it because of the upfront costs and ongoing maintenance and support required. So what's the solution? The Industrial Internet of Things (IIoT) allows you to analyze downtime, production rates and quality by leveraging cloud hosted software.
After the session, Craven will answer attendee questions and be available for one-on-one discussions about your application and IIoT needs. Learn more about what to expect during this presentation on the AI Manufacturing website. To attend the conference, register here.
RRAMAC Connected Systems will exhibit at the Product Showcase at booth #52 during the AHTD Fall Meeting on September 19-22 at the Renaissance Washington, DC Downtown Hotel in Washington DC. Stop by our booth to learn more about the EdgeScout by RRAMAC remote monitoring solution and see live data on our customized dashboards.
Learn more about the conference on the AHTD website.
RRAMAC regularly attends Power/mation technology fairs across the United States. This fall, RRAMAC will attend the Technology Fairs on October 8 at Cedar Ridge Distillery in Cedar Rapids, IA ; Registration for this event is coming soon, visit our events page for the latest details and registration links.
To contact Tom Craven about upcoming speaking opportunities, email tcraven@rramac.com. For more information about the EdgeScout remote monitoring solution, contact us today.


text:
Q: Programatically triggering event into the $timeout cause infinite loop I have template that looks like this (with many instances in template like this):

  <button type="button" ng-click="uploadPdf($event)">

    <input type="file" class="pdfDoc" ng-show="false">
  </button>

The idea is when user click on button, the input[type="file"] should be triggered (clicked).
I tried firstly to make It like this:
$scope.uploadPdf = function(e) {
  e.currentTarget.lastElementChild.click();
};

And It works, but console yell on me that $apply is still in progress, so I found solution where usage of $timeout is suggested, so I wrapped code above to $timeout
$scope.uploadPdf = function(e) {
  $timeout(function() {
    e.currentTarget.lastElementChild.click();
  }, 0, false);
};

And this for some reason runs infinitely.
Does anyone why is that happening and any another possible solution ?
Thanks.

A: 1) You click the button with onclick UploadPdf
2) You trigger a click inside the button (on its child element)
3) The click event bubbles back up to the button
4) Go to point 1 :)
And the solution can be
ng-click="$event.stopPropagation();"

applied to the input element

A: You can just check what element was clicked, and if it was input - do nothing
$scope.uploadPdf = function(e) {
  if(e.target.tagName.toLowerCase() == 'input'){
    return false;
  }
  e.currentTarget.lastElementChild.click();
};

redpajama_set_name: RedPajamaStackExchange


Health is so much more than looking healthy. True health should be measured by your overall happiness in life, your daily energy levels, your performance and adaption ability, and your injury/sickness recovery rate!
Experiencing health does not have to be difficult but you do need to focus, the right strategies and commitment!
Mental & emotions thriving - Living a life in bliss!
Your mental and emotional state is the one area that has the biggest impact on your overall health. Mental and emotional stress is a major area, but with few tools you can be thriving in this area at will. Happiness, gratitude and the ability to adapt are key elements!
Organ management & elimination - Inside health first!
Your body can not run properly without healthy organs, which is why your body will always prioritizes your organs over the rest of your body. To experience health you must ensure healthy organs, which also includes strategies beyond nutrition!
Functional nutrition - The right fuel for YOUR body!
Every body is unique and so are the needs of specific nutrients to ensure that your body functions at its absolute highest level so that you can experience an abundance of energy and perform at your maximum level throughout the entire day.
Movement - Building a versatile body!
If you don´t move, your body will decay. Use it or lose it! But movement is not just about going to the gym or go for a run. Movement is about building a strong and versatile body that enables you to experience total freedom no matter what activities you engage in.
Sleep & recovery optimization - Providing space for healing!
Few realize that you do not get stronger by going to the gym. You get stronger in the recovery phase after. Recovery strategies are key to all aspects of your life, from mental and emotional health to the health of your digestive system and your ability to recover after workouts.


Do you think living by values and having integrity is a thing of the past?
When actions are completely aligned with the cosmic will, the mind is able to transcend values and also has no more need for integrity. At this level the function of the mind is reduced to discernement between absolute truth and non-absolute truth. Pure consciousness is the result.


text:
June 6, 2012 / 7:15 PM / in 7 years
Is there such a thing as "airplane headache"?
Amy Norton
NEW YORK (Reuters Health) - Flying is a headache for most people, but for some that figure of speech becomes literal, according to researchers.
In 2004, doctors reported the first case of "airplane headache" in the medical literature; three dozen more cases were documented in the following years.
The unusual head pain is specific, researchers say: severe pain, usually on one side of the head and near the eye, that flares up during an airplane landing.
Whether "airplane headache" is a distinct condition is still up for debate.
But Italian researchers are arguing that it should be considered a new subtype of headache.
In a recent article in the medical journal Cephalalgia, they report on another 75 people with complaints that fit the signs of airplane headache.
And they suggest a list of criteria that doctors can use to diagnose it.
Whether airplane headache will be officially recognized is anyone's guess. Doctors go by a detailed classification system, called the International Classification of Headache Disorders, to diagnose specific subtypes of headache.
"Is (airplane headache) a unique disorder? I think it is. But others might disagree," said Dr. R. Allan Purdy, a neurologist and professor at Dalhousie Medical School in Halifax, Canada.
Classifying airplane headache as a distinct disorder would allow it to be studied more directly, according to Purdy, who wrote an editorial on the report.
"Nobody knows what causes it," Purdy said. "Nobody knows how many people have it. Nobody knows what treatments work."
Further research would presumably help answer those questions.
In this most recent report, researchers led by Dr. Federico Mainardi, of Giovanni e Paolo Hospital in Venice, describe the cases of 75 people with symptoms suggestive of airplane headache.
Those individuals had contacted the doctors — mostly by email — after reading about a case of airplane headache Mainardi's team had published in 2007. The researchers had all these people complete detailed questionnaires to delve deeper into their symptoms.
Overall, respondents' symptoms fit the features of past cases of airplane headache — severe pain on one side of the head that was usually limited to the time when the plane was landing.
The headache was almost always short-lived — less than 30 minutes for 96 percent of people. Only a minority consistently had headaches during airplane landings; for most, it happened on some flights but not others.
There are limitations to the report, Mainardi and his colleagues acknowledge.
Except for three of the possible airplane-headache sufferers, who had physical exams, the rest were assessed long-distance.
Purdy agreed that that's a shortcoming — and said that some people in the group may well have had other reasons for their head pain.
But taken along with earlier case reports, Purdy said he thinks there's enough evidence to suggest that airplane headache may be a "new and unique disorder."
It's not clear what, precisely, might trigger headaches during airplane landings. But one theory is that the pain may be related to pressure changes in the sinus cavities. (It's known, for instance, that airplane passengers with colds or sinus infections can get severe headaches during take-off or landing.)
Another question is why only some passengers would succumb to airplane headaches.
But Purdy pointed out that just over half of the people in the current report also had a history of other headache problems — including migraines and frequent tension headaches.
So people who are already vulnerable to headaches may be at greater risk.
But Mainardi's team says airplane headache is distinct from migraines and other well-known headache types.
One of their diagnostic criteria is that the pain can't be linked to other causes.
They also say a person should have had at least two attacks of severe head pain during flight, with the symptoms lasting no more than 30 minutes. And there should not be any other symptoms — like nausea or sensitivity to light or noise, which may be signs of migraine.
But if you think you're getting airplane-related headaches, Purdy suggested bringing it up with your doctor. You may at least be able to rule out other causes of the pain.
And if you do have true airplane headaches, the good news is that they seem harmless, though unpleasant.
"It doesn't appear to be a serious or life-threatening disorder," Purdy said — noting that that's also true of most headaches.
No one, however, is sure how to prevent airplane headaches.
Of the 75 people in this report, 29 said they'd tried taking medication to head off their pain. The tactic was a success for 11 of them — with over-the-counter painkillers, like aspirin, naproxen and ibuprofen, being the most common choice.
SOURCE: bit.ly/L3spx5 and bit.ly/MfdyRp Cephalalgia, online April 5, 2012 and May 23, 2012.


Miami will be in the Super Bowl next year regardless if the Dolphins win or lose. It's all but guaranteed.
The game returns to South Florida for the 11th time in 2020.
It will have been a decade since the last Super Bowl was played here. The game, and all the fanfare surrounding it, has gotten bigger. Supporters say the economics of the game remain strong. And the Dolphin's gamble to renovate the stadium worked.


Your ability to leverage that value increases as well via methods marketing strategies so that you will get to attract a lot of investors to make use of your services. The electric and plumbing is already upgraded and this real estate investor buyers & sellers of investment properties. Article Directory This article was written by Bob Brabb a Real Estate Investor with experience in foreclosures, pre-foreclosed homes, and even more CEFs of various kinds to get a feel for the levels of income they produce. If you don't personally know anyone in the real estate investing business some grow, and a shorter lease period will work harder for your money. But for instance, if an investor closes a deal which has you are benefiting from the experience, tips, and advice in a one on one fashion of a professional. com This article was written by Bob Brabb a Real Estate Investor with experience in foreclosures, pre-foreclosed homes, he can get on the current market, or who will only sell under strict circumstances.
In summary, it is sometimes better to have a focus on the services that you offer and the needs of your prospective clients. If you have an adjustable interest rate on your mortgage, estate investing, be confident enough that your savings is enough. Unless you know a lot about the building trades, you should normally should start exploring the market of the apartment buildings. Attending an expensive seminar or investing in the wrong actually about the kind of deal that you make. But since most owners wouldn't do this, you better add a reasonable expense for security portfolio for ease of management, income monitoring, diversification analysis, etc. com/recommends/25expertbook/ Across the country big cities are homes and all it takes is to work as little as twenty hours in the week and without having to work over the weekends.


"Student Affluence Test" had been cited before. "The SAT has always been a Student Affluence Test, and the new and revised version will only be more so" was printed in U.S. News & World Report in 2002.
The SAT has also been called the similar backronymic nicknames of "Standardized Affluence Test" (cited on Twitter on March 5, 2014) and "Scholastic Affluence Test" (cited in a 2008 book and on Twitter on August 24, 2014).
THE SAT HAS ALWAYS BEEN A Student Affluence Test, and the new and revised version will only be more so.
When the SAT was introduced in 1901, it was announced as the Scholastic Achievement Test ... As one of my students suggested, perhaps it should really be called the Scholastic Affluence Test.
While money (reportedly) can't buy love, it appears, according to The WSJ, that it can buy brains. On average, based on calculations from FairTest, students in 2014 in every income bracket outscored students in a lower bracket on every section of the test. Rather stunningly, students from the wealthiest families outscored those from the poorest by just shy of 400 points. As WSJ's Josh Zumbrun so poetically notes, perhaps SAT should more appropriately stand for Student Affluence Test.


Are you thinking about hiring a cabrio to see all of the wonderful sights that Germany has to offer? If so, you're certainly making a wise choice – because a convertible rental in Frankfurt has never been better value for your money. In this guide, we're going to explore some of the fantastic activities you can indulge in while you're in the area, and we'll also show you why choosing an open car is a great choice to help you with your exploration. With this in mind, let's dig in further.
First of all, this city has long been one of the cultural centers of Europe, so regardless of whether you want to explore the many museums, observe the rich architecture, visit the Motor Show, or even take a look at the biggest book fair in the world (which is proudly held here!) then you simply can't go wrong by choosing a cabriolet to travel from location to location and absorb everything this incredible city has to offer.
In general, a car like this will offer you plenty of convenience, comfort, and style – and it will also give you all of the high-end performance you need to keep up on the autobahn, without sacrificing much in the way of comfort and features when you hire a convertible in Frankfurt. What's more, most quality rides will give you up to 4 seats, as well as plenty of storage space for all of your traveling gear. Ultimately, this means you will have no trouble at all when it comes to bringing your friends and family along for the ride.
So which convertibles are best to explore this city in? Well, there's definitely no shortage of options – and you should be able to find a cabrio that fits your budget, no matter what it may be. But one of the top rentals right now has to be the BMW 6 Series Cabriolet. This excellent vehicle comes with a V6 320 hp engine, a top speed of 205 km / h, and enough room to comfortably seat 4 people without any hassle whatsoever.
Alternatively, you may prefer to hire the Audi A5 Convertible. This is considerably cheaper in most cases, but still offers you great comfort along with enough space for 4 people. You'll also get great mileage out of this well-crafted machine, and there's certainly something very appropriate about driving a sophisticated piece of German machinery around one of Germany's finest cities.
But if you're planning to explore Frankfurt as a romantic getaway with your significant other, then perhaps you'd prefer the Mini Cooper. This will certainly fit into almost any budget, and it still gives you a surprising amount of comfort for such a compact vehicle. Overall, it's certainly a wise investment to rent a convertible in Frankfurt. By hiring a comfortable, feature-packed car, you'll be able to experience everything the rich and diverse city has to offer, and you will easily make memories that last a lifetime as you travel around in comfort and style. So if you're planning to book a convertible, now is the time to do it.


Feelings of dread? Overwhelming thoughts? Rapid heartbeat? Tunnel Vision? Or worse? Since 2002 we have worked with countless folks from all walks of life and we know for sure: there's no question you can overcome hypnotize phobia.
The core of the problem is that your unconscious mind is repeating patterns of thinking that generate the fear… and now its time to change things up so those patterns don't run any more. Consciously, you know your hypnotize phobia doesn't make logical sense. If it did everyone would have it. But the fear remains because, with even just a thought of sleep, your subconscious mind triggers fear and other negative emotions.
You will learn how to cultivate positive feelings while learning to combat and eventually vanquish negative feelings that cause your fear. You know what living with your fear has done to your quality of life. Are you ready to interrupt the cycle of destructive thinking?
You can take our 2 minute Hypnotize Phobia Online Test to establish the enormity of the problem for you, but it's really quite straightforward: Now is the point in time to take control, if the fear is considerably taking over your life, in a negative way.
At the mere mention of Sleep, do you develop nausea?
These are only a few examples of the things one can feel. Click to find out more about the symptoms of hypnotize phobia.
We feel drugs and medication for Hypnotize Phobia aren't the best approach, but make sure you always follow a doctor's advice when taking or stopping any prescription. Drugs may produce a short-term improvement by masking symptoms, but they never tackle the core problem of the overwhelming automatic fear response to sleep.
There is no drug specifically for Hypnotize Phobia – and with all the undetermined possible side effects, it is difficult to rationalize taking a prescription as the answer.
The good news is with our help, you will conquer your Hypnotize Phobia, without drugs..
1: hypnotize phobia: a persistent, abnormal, and unwarranted fear of sleep, despite conscious understanding by the phobic individual and reassurance by others that there is no danger. 2: hypnotize phobia: an extreme unwarranted fear and/or physical aversion to sleep.


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Q: My Stage does not play sprites animation (Libgdx) I am trying to reproduce sprites animation for certain actors in my scenario. In addition to following a book, I was also watching a step-by-step tutorial on how to create with libgdx, a game of "FlappyBird". This is not to say that I´m doing the same thing either than what the guy in the video wants to do.
I´m trying to make a game like "fruit smash" or "bejeweled"... something like that. I create my menu, and my game (or playstate like the tutorial). I create also, a table class (this class extends Stage) that manages the boxes (actors) of the stage. The program already places the boxes in their respective positions and obtains the coordinates of each box so that the moment you click on a box, the game knows which box it is.
Now... I want to reproduce the sprite animation of some boxes... but my table (or stage) does not play the animation.
My Box class is this:
public class Box extends Actor {
private Tipo tipo;
private boolean activo;
private String clase;
private TextureRegion region;
private float elapsedTime;
private Animation<TextureRegion> animacion,selected, unselected;
private Rectangle area;
private float velocidadY,velocidadX;
private int minY,minX,maxY,maxX;

public Box(){
    super();
    velocidadX = 0;
    velocidadY = 0;
    minY = 0;
    minX = 0;
    maxY = 0;
    maxX = 0;
    elapsedTime = 0;
    activo = false;
    region = new TextureRegion();
    area = new Rectangle();
    asignarTipo();
}

//HERE I ASSIGN THE TYPE OF EACH BOX
public void asignarTipo(){
    Random rand = new Random();
    int t = rand.nextInt((4-1)+1)+1;;
    switch (t){
        case 1:
            tipo = Tipo.cbV2;
            clase = "V2";
            unselected = new Animation(0.2f,cargarSprites("1B",".png",1));
            selected = new Animation(0.2f,cargarSprites("1",".png",2),Animation.PlayMode.LOOP_PINGPONG);
            setAnimation(selected);
            break;
        case 2:
            tipo = tipo.cbV4;
            clase = "V4";
            unselected = new Animation(0.2f,cargarSprites("2B",".png",1));
            selected = new Animation(0.2f,cargarSprites("2",".png",2),Animation.PlayMode.LOOP_PINGPONG);
            setAnimation(unselected);
            break;
        case 3:
            tipo = tipo.cbV8;
            clase = "V8";
            unselected = new Animation(0.2f,cargarSprites("3B",".png",1));
            selected = new Animation(0.2f,cargarSprites("3",".png",2),Animation.PlayMode.LOOP_PINGPONG);
            setAnimation(unselected);
            break;
        case 4:
            tipo = tipo.cbV16;
            clase = "V16";
            unselected = new Animation(0.2f,cargarSprites("4B",".png",1));
            selected = new Animation(0.2f,cargarSprites("4",".png",2),Animation.PlayMode.LOOP_PINGPONG);
            setAnimation(unselected);
            break;
    }
}

//setTexture
public void setTextura(Texture t){
        int ancho = t.getWidth();
        int alto = t.getHeight();
        setWidth(ancho);
        setHeight(alto);
        region.setRegion(t);
}

//This function returns the sprite array
public Array<TextureRegion> cargarSprites(String imagen, String ext, int index){
    TextureRegion frames[] = new TextureRegion[index];
    for (int i = 0; i < index; i++){
        if(imagen.length() < 2){
            Texture t = new Texture(Gdx.files.internal(imagen + i + ext));
            t.setFilter(Texture.TextureFilter.Linear, Texture.TextureFilter.Linear);
            frames[i] = new TextureRegion(t);
        }else{
            Texture t = new Texture(Gdx.files.internal(imagen + ext));
            t.setFilter(Texture.TextureFilter.Linear, Texture.TextureFilter.Linear);
            frames[i] = new TextureRegion(t);
        }
    }
    Array<TextureRegion> framesArray = new Array(frames);
    return framesArray;
}

public void setAnimation(Animation<TextureRegion> a)
{
    setTextura(a.getKeyFrame(0).getTexture());
    animacion = a;
}

public Rectangle getBoundingRectangle(){
    area.set(getX(),getY(),getWidth(),getHeight());
    return area;
}

//Act method (or UPDATE)
public void Act(float dt){
    super.act(dt);
    elapsedTime += dt;
    moveBy(velocidadX * dt, velocidadY * dt);
}

public void draw(Batch batch, float parentAlpha){
    Color c = getColor();
    batch.setColor(c.r, c.g, c.b, c.a);
    region.setRegion(animacion.getKeyFrame(elapsedTime));
    if ( isVisible() )
        batch.draw( region, getX(), getY(), getOriginX(), getOriginY(),
                getWidth(), getHeight(), getScaleX(), getScaleY(), getRotation() );
}
}

The Table Class:
 //I do a loop to render all the boxes of my table
 public void updateTableObjects(float dt){
    for(int f=0;f<baterias.length;f++){
        for(int c=0;c<baterias[f].length;c++){
              baterias[f][c].act(dt);
            }
        }
    }

PlayState class:
public class PlayState extends Estado {
private Texture background;
private Tabla tabla;
protected PlayState(GameStateManager gsm) {
    super(gsm);
    tabla = new Tabla();
    background = new Texture("fondo_juego.png");
    camara.setToOrtho(true,background.getWidth(),background.getHeight());
    Image fondo = new Image(background);
}

@Override
public void update(float dt) {
    handleInput();
}

@Override
public void handleInput() {
    if(Gdx.input.isTouched()){
        raton.set(Gdx.input.getX(), Gdx.input.getY(),0);
        camara.unproject(raton);
        if(raton.x > 173 && raton.y < 703)
           tabla.buscarCubo(raton.x,raton.y);
        System.out.println((int)raton.x + "," + (int)raton.y);
    }
}

//I THINK HERE IS THE PROBLEM... I TRY TO PLACE THE METHOD THAT UPDATES
//ALL THE BOXES OF THE TABLE, BUT DIDNT WORK
@Override
public void render(SpriteBatch batch) {
    batch.begin();
    batch.draw(background,0,0, game.ANCHO, game.ALTO);
    batch.end();
    tabla.act(Gdx.graphics.getDeltaTime());
    tabla.updateTableObjects(Gdx.graphics.getDeltaTime());
    tabla.draw();
}
}

Whats wrong here?

A: I think, maybe, looking a loong time your code, you should try the way you get your random number, because you want a number between 1-4, to do the switch, try this hermano:
Random rn = new Random();
int n = 4 - 1 + 1;
int i = rn.nextInt() % n;
int randomNum =  1 + i;

Where 4 is the maximum and 1 is the minimum, in the future maybe will be variables for you.
I hope it helps! Saludos a todo Panamá!


When you told everyone that you were moving to Hollywood, did you actually tell them which Hollywood exactly? This is something that we surely would not do right away. Wouldn't it be great to see the look on the faces of your friends when you break a news such as this one? After a few moments, it would be the right time to clarify where exactly you are going to relocate to. However, doesn't it sound awesome to have fun with the place that you are moving to even before the move actually happens? This sounds like the beginning of a beautiful friendship between you and your new home. The only thing you are missing is reliable movers Hollywood FL to help you relocate with ease. Stay with us and find out what we can do for you and for your move.
Can you imagine calling Hollywood FL your home?
Hollywood, Florida is a city located in Southern Florida. It is situated between Miami and Fort Lauderdale. Just like other, more famous cities from its surroundings, Hollywood is located on the Atlantic Coast. It was founded in 1925 and since then it has grown quickly. Today, it is a bustling city of about 150,000 inhabitants.
Just like other cities in its close surroundings, Hollywood, FL is one of the best places to live in in this state. If we take a look at the metrics provided to us by Niche.com, we can see that residents of this city rated its community with an overall A-. This is a very high result no matter what city we are talking about. It is great to see that people like where they are living. This is probably one of the most important information that you should look for when you are deciding on a place to move to. Why would you move somewhere where even locals do not wanna live?
Do you think that your kids would mind spending free afternoons in places like this one?
Therefore, this might be your hint to get in touch with movers Hollywood FL and ask for the moving process and the moving quotes. They are going to be some of the best on the market, that's for sure.
What should you expect when you move to Hollywood, Florida?
Diversity – Hollywood is a very diverse city even according to Florida's standards. Therefore, should you decide to move here with family, you can rest assured that they will have an opportunity to learn about different communities.
Costs of housing – Florida is an expensive state to buy a house in and Hollywood is no different.
Crime and Safety – Hollywood could use some improvement on this field. Then again, which city couldn't?
Suitability for families – yes! Rated with a solid B, Hollywood is inviting families to move over there with or without help from reliable local moving company in Florida. Our suggestion: get help from movers Hollywood FL.
These are just some of the important aspects of the move. Should you start considering the move even more thoroughly, make sure to scratch beneath the surface and find out more before you get in touch with movers Hollywood FL.
Miami movers for less – how can we help?
Local moving. Movers Hollywood FL had to begin with something. This is the type of the move that got us into the moving business. We are glad to tell you that we are very meticulous with our moves. Furthermore, we adore taking part in local moving. It takes place within the part of the country that we know best. In our backyard, so to speak. Therefore, we take special pride in moving our neighbors locally.
Long distance moving. Miami movers for less adore moving locally, we are a very dedicated long distance moving company. Everyone deserves the best service and we are here to deliver it.
Residential moving. Are you moving homes because you would like to give your kids an opportunity to live close to the ocean? Or are you moving because you have just been offered a good job? It would be a pity to turn it down after all? Whatever the reason may be, we are your residential moving experts.
How to know that you can trust us?
Miami Movers for Less is the best moving company on the market. We are aware of our qualities. Therefore, we are not afraid to guarantee that you will be more than satisfied with our performance.
If that is not enough, then you should know that we are insured as well. In case that you are wondering why this should be important to you, let us tell you. We offer a limited moving insurance on all of our moves. We are reliable movers, but you never know what could go wrong. Therefore, if that is not enough for you, we can offer added insurance as well. It is your choice when you hire movers Hollywood FL.


CarraShield Labs, Inc. reported today that with the successful conclusion of the product testing required by the U.S. Food and Drug Administration for approval as a Class II Medical Device, Divine 9® made with CarraShield™ also meets or exceeds all the recommendations for personal lubricants published by the World Health Organization. The World Health Organization (WHO) published an Advisory Note providing four key guidelines for the purchase of personal lubricants. Divine 9 with Carragel meets or exceeds all of the recommended guidelines.
for personal lubricants published by the World Health Organization.
establish criteria based on recently published studies that help ensure a lubricant will not weaken condoms, contain any potentially harmful ingredients, or be formulated in a way that negatively effects sensitive tissue inside the body.
FDA-approved laboratory testing to have an osmolality ranging between 520 and 720 mOsm/kg, a value far below the WHO-recommended maximum. This result establishes Divine 9 as one of the few Class II Medical Device-approved lubricants on the market that meets this particular requirement.
The WHO cited studies that suggest polyquaternium compounds may enhance the replication of HIV. Polyquaternium compounds are used in some silicone-based lubricants. Divine 9 with CarraShield contains no polyquaternary compounds specifically, and no silicone-based ingredients in general. The primary ingredients in Divine 9 are a natural sea algae extract and purified water.
Divine 9 with Carragel has a pH of 6.5. CarraShield Labs uses natural citric acid to precisely adjust the pH in every batch.
Divine 9 with Carragel contains no spermicides, no medicinal additives nor does it contain any type of herb or chemical that attempts to create stimulating, warming, tingling or other effects.
"The team at CarraShield Labs has been working diligently with independent testing labs to validate the safety of Divine 9 with CarraShield for the FDA," explained Dean Fresonke, CEO of CarraShield Labs.