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Comprehensive analysis of PD-L1 expression in glioblastoma multiforme
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Dieter Henrik Heiland1,4, Gerrit Haaker1,4, Daniel Delev1,4, Bianca Mercas1,4, Waseem Masalha1,4, Sabrina Heynckes1,4, Annette Gäbelein1,4, Dietmar Pfeifer2,4, Maria Stella Carro1,4, Astrid Weyerbrock1,4, Marco Prinz3,4,5 and Oliver Schnell1,4
1Department of Neurosurgery, Medical Center - University of Freiburg, Baden-Württemberg, Germany
2Department of Hematology, Oncology and Stem Cell Transplantation, Medical Center - University of Freiburg, Baden-Württemberg, Germany
3Institute of Neuropathology, Medical Center - University of Freiburg, Baden-Württemberg, Germany
4Faculty of Medicine, University of Freiburg, Baden-Württemberg, Germany
5BIOSS Centre for Biological Signalling Studies, University of Freiburg, Baden-Württemberg, Germany
Dieter Henrik Heiland, email: [email protected]
Keywords: glioblastoma multiforme, PD-L1, WGCNA, integrative analysis, immunotherapy
Received: September 09, 2016 Accepted: January 10, 2017 Published: February 02, 2017
Glioblastoma multiforme are highly malignant brain tumours with frequent genetic and epigenetic alterations. The poor clinical outcome of these tumours necessitates the development of new treatment options. Immunotherapies for glioblastoma multiforme including PD1/PD-L1 inhibition are currently tested in ongoing clinical trials. The purpose of this study was to investigate the molecular background of PD-L1 expression in glioblastoma multiforme and to find associated pathway activation and genetic alterations. We show that PD-L1 is up-regulated in IDH1/2 wildtype glioblastoma multiforme compared to lower-grade gliomas. In addition, a strong association of PD-L1 with the mesenchymal expression subgroup was observed. Consistent with that, NF1 mutation and corresponding activation of the MAPK pathway was strongly connected to PD-L1 expression. Our findings may explain different response to PD-L1 inhibition of patients in ongoing trials and may help to select patients that may profit of immunotherapy in the future.
Glioblastoma multiforme (WHO grade IV) is the most common type of brain tumours, which is characterised by poor clinical outcome and short survival time, rarely longer than 14 months . Extensive efforts have been made to develop new treatment strategies during the last decades, without improving the poor clinical course of glioblastoma multiforme [2, 3]. Until now, the “gold standard” in glioblastoma multiforme therapy remains surgery plus adjuvant combined chemoradiotherapy introduced by Stupp et al., 2005 .
Glioblastoma multiforme contains a high frequency of genetic and epigenetic alterations with numerous potentially produced neoantigens , which are recognized by the immune system and support T-cell based immune response . So far, new treatment strategies are investigated for clinical testing. Those treatments target the programmed death ligand 1 (PD-L1) or the programmed death ligand protein 1 (PD-1) receptor and attempt to re-activate the immune system. Currently, PD-1/PD-L1 therapeutic antibodies are investigated for a large variety of different cancer types. For instance, in small-cell lung carcinoma and melanoma, PD-1 inhibitors were successfully tested in clinical trials and could significantly improve the overall survival [6–8].
The programmed death ligand 1 (PD-L1 or CD274) plays a major role in preventing the immune response in many cancer types [6, 8, 9]. This mechanism of immune system escape is described as “tumour immunity” [10–12] and represents as a hallmark of cancer biology. Recent studies have reported a strong occurrence of PD-L1 in glioblastoma, which is targetable by prospective immunotherapies . The underling regulation mechanism is not well described and rarely explored. Bloch et al. 2013 reported a regulation of PD-L1 by tumour associated macrophages and related Il-10 signalling . Some years ago, Parsa et al., 2006 showed a relationship between loss of PTEN and increased expression of PD-L1 in glioblastoma multiforme . However, the association was not observed in following studies as the study by Berghoff et al., 2015 . Glioblastoma multiforme are tumours with an aggressive local growth pattern, including strong migration, proliferation and invasion of normal brain . Invasive parts of glioblastoma multiforme were associated with massive metabolic alterations within hypoxia and HIF1A up-regulation which lead to an increased expression of PD-L1 .
Until now, genetic alterations and specific tumour related pathways that support PD-L1 depended “immune escape” in glioblastoma multiforme are rarely explored.
In other cancer types, PD-L1 expression and its clinical impact are more frequently investigated. Madore et al., described a strong association between PD-L1 expression and mutational load in melanoma . In addition, an up-regulation of immune response pathways was found in PD-L1 positive melanoma . Another study investigated the regulation mechanism of PD-L1 by activation of PI(3)-kinase and phospho-S6-kinase in breast cancer . Chen et al., described different mechanism of PD-L1 regulation as PI(3)Kinase and MAPK pathway . Also, an association of the hypoxic pathway and STAT3 transcriptional regulation was found [16, 20].
The study reported here, combines data from both the TCGA Data Platform and local tumour-bank in order to perform an integrative analysis of PD-L1 expression in glioblastoma multiforme. These results may improve further diagnostics and identify glioblastoma multiforme subgroups that could profit from PD-L1 targeted therapy and may explain different clinical courses of patients in ongoing clinical trials.
PD-L1 expression/methylation patterns in glioma subgroups
PD-L1 was differently expressed according to the tumour grade (Figure 1A). WHO grade IV tumours, showed significantly stronger expression compared to lower-grade gliomas (WHOII-WHOIII p>0.05, WHOII/WHOIII-WHOIV p<0.05). Moreover, analysis of PD-L1 expression in different glioblastoma multiforme subgroups (Verhaak et al., 2010 ) revealed an increased expression of PD-L1 in mesenchymal tumour samples (p<0.05) (Figure 1B), whereas the lowest PD-L1 expression was found in proneural tumours.
Figure 1: A. A boxplot PD-L1 expression in patients with WHO grad II, III and glioblastoma multiforme (WHO grade IV). B. A boxplot of PD-L1 expression in all expression subclasses define by Verhaak et al., 2010 of high-grade gliomas (glioblastoma multiforme). C-D. Survival Analysis of the TCGA database and Freiburg cohort. PD-L1 high vs low was determinate by the mean expression +/- standard deviation. Patients of the Freiburg cohort with outstanding events were censored and marked by a cross. *** p<0.001, ** p=0.01, *p<0.05.
Survival analysis of the TCGA patients showed no differences in the overall survival (OS) of patients with high PD-L1 expression (define as: mean PD-L1 expression plus standard deviation, mean OS: 427 days) or low PD-L1 expression (define as: mean PD-L1 expression minus standard deviation, mean OS: 426 days) with a Hazard ratio (HR) of 0.98 (p>0.05), Figure 1C. Similar results were found in the Freiburg cohort with a mean OS of 595 day in the PD-L1-high group vs. 611 days in patients with low PD-L1 expression (HR 0.94 p>0.05), Figure 1D.
Brad et al., described three distinct molecular subtypes in lower-grade gliomas (WHO II-III) . Analysis of these subgroups (IDH1/2 mutation, IDH1/2 mutation+1p19q Co-deletion, IDH1/2 wild-type) revealed a strong PD-L1 expression in the IDH1/2 wild-type samples (Figure 2A). The methylom of patients with IDH1/2 mutation presented enormous epigenetic alterations, described as the hypermethylated phenotype (G-CIMP) . Those alterations may influence the PD-L1 expression by epigenetic silencing. In an additional methylation analysis of the PD-L1 promoter region, a hypermethylation in IDH1/2 mutated patients was found (Figure 1C, 1D), possibly explaining the downregulation of PD-L1 expression in proneural glioblastoma, which are partly G-CIMP (Figure 1D). Similar findings were investigated in glioblastoma multiforme patients with and without IDH-mutation (p<0.01).
Figure 2: A. A boxplot of PD-L1 expression in lower-grade gliomas and its molecular subgroups. B. A boxplot of PD-L1 expression in IDH mutated and non-mutated glioblastoma multiforme. D. A boxplot of PD-L1 promoter methylation in IDH1/2 wild-type and mutated patients. C. Functional analysis of methylation/expression correlation of different CpG sides of the PD-L1 promoter. The strongest negative correlation was found in the cg19724470. Significant values were corrected by false-discover rate (FDR). (D) This CpG side was used to validate the mean methylation of IDH mutated vs. non-mutated patients. *** p<0.001, ** p=0.01, *p<0.05.
Copy-number-variations (CNV) and mutational analysis
Whole-exome sequencing data of 264 patients derived from the TCGA database were analysed. 19 mutations with high occurrence were selected as shown in Figure 3A. Expression of PD-L1 was not directly correlated to the total mutation load of each patient (R=0.2 p>0.05). However, after performing a logistic regression analysis, NF1 mutation was identified as a predictor of increased PD-L1 expression. Other very frequently occurring mutations like PTEN (p>0.05), EGFR (p>0.05) or TP53 (p>0.05) did not correlate with PD-L1 expression. However, the effect of NF1 alteration on PD-L1 expression was not exclusively for NF1 mutation, also patients with NF1 deletions showed increased expression of PD-L1 (p<0.01 for homozygote and heterozygote deletions). These findings could be confirmed in a validation cohort (Freiburg) (p<0.01 heterozygote deletions) (Figure 3B). IDH 1/2 mutation was found negatively associated with PD-L1 expression (as described above). This result is in line with the above-described PD-L1 expression in the IDH1/2 mutated subgroups.
Figure 3: A. Co-mutation plot of highly frequent mutations in glioblastoma multiforme. Mutational frequencies were given in a bar plot beside the co-mutation plot. General mutation load as total numbers of mutations in each patient was presented in the top barplot. PD-L1 expression was illustrated in a barplot in the panel below. Patients with missing expression values were marked by *. B. Boxplots of PD-L1 expression (TCGA) in NF1 wild-type and mutated/deleted samples were given in the left panel. The validation cohort of Freiburg patients showed similar expression differences (right panel). C. Protein-level of p-c-RAF and p-S6 in mutated/deleted and wild- type samples were illustrated. D. Weighted Proteome Network Analysis of the proteome data investigated protein module 1 as highly correlated to PD-L1 expression level. E. Created network analysis of protein module 1 based on intramodule connectivity, derived from WPNA.
Weighted proteome correlation network analysis (WPCNA)
Proteome data of the TCGA database were used as input for a network analysis. The purpose of the analysis was to describe a PD-L1 regulatory network at protein level and connected proteins were summarised in modules by WPNA (Figure 3D). Next, the modules that showed an association with PD-L1 expression were analysed by its intramodule-connectivity in correlation to PD-L1 expression. Protein module 1 (PM1) was identified as highly correlated to PD-L1 expression (R=0.32 p=2.0E-5). A network was calculated based on the intramodule connectivity of each protein in PM1 (Figure 3E). The WPNA of proteome data revealed an association of the RAF-RAS-MAPK pathway with PD-L1 expression. High protein-level of phospho-c-RAF (p<0.05) and phospho-S6 (p<0.05) were associated with NF1 loss-of-function (Figure 3C) and resulted in an increase of PD-L1 expression (Figure 3B, 3C). An additional illustration of associated protein activation downstream of the NF1 pathway is shown in the supplementary Figure 1. Immunostainings of 16 patients (8 Patients with heterozygote NF1-deletion 8 patients without NF1 alteration) of the Freiburg cohort were performed to validate the coherence of MAPK-pathway activation and PD-L1 expression. A significant differences between patients with/without a NF1 loss-of-function. Additionally, p-MAPK14 (p-p38) as a maker of the MAPK-pathway activation was positively correlated (r=0.6, p<0.01) to PD-L1 expression (Figure 4D). P-JNK was also increased in NF-deleted patients, while p-AKT more increased in wild-type patients Figure 4E, 4F.
Figure 4: A-C. Immunostaining of PD-L1 and phospho-MAPK14 in two patients with NF1 deletion (A) and two patient without NF1 alteration (C). Additionally, six independent fields were quantified by mean signal intensity by ImageJ, normalized and illustrated in a boxplot. D. Correlation analysis of protein-level of PD-L1 and p-MAPK showed a strong positive correlation (r=0.6 p<0.01). E-F. Immunostaining of phospho-JNK and phospho-AKT in a patient with NF1 deletion (upper panel) and one patient without NF1 alteration (lower panel). Additionally, quantification was given in the right boxplot (F). *** p<0.001, ** p=0.01, *p<0.05.
Weighted gene co-expression network analysis (WGCNA)
The WGCNA identified several modules (Figure 5A) that were clustered based on its module eigengene vector (Figure 5B). Two modules were addressed in a cluster together with PD-L1 expression levels. The correlation of each module and its association to PD-L1 was validated by direct correlation analysis of PD-L1 expression and intramodule connectivity.
Figure 5: A. Weighted gene co-expression network analysis of the whole transcriptome data. B. A cluster of clusters analysis presented connected expression modules. Expression module 1 and 2 were contained in one cluster. C. Detailed cluster branches of expression module 1 and 2 were illustrated. Corresponding correlation heatmaps of PD-L1 expression and module-contained genes are given in the panel below the branches. Scatterplots of intramodule connectivity (KME) confirmed the strong correlation of PD-L1 expression and the expression modules. D. Gene Set Enrichment Analysis of expression module 1 identified INFγ related genes (upper panel) and immune response (bottom panel). E. Created network analysis of expression module 1/2 based on intramodule connectivity, derived from WGCNA. Size and colours indicate the intensity of intramodule connectivity.
Expression module 1 (Figure 5C, yellow) was significantly correlated with PD-L1 expression (R=0.54 p=2.2E-40). Permutation based pre-ranked GSEA was performed and revealed a significant enrichment of mesenchymal subgroup genes (pFWER<0.01) (data not shown). Expression module 2 (Figure 5C, violet) was also correlated with PD-L1 expression (R=0.5 p=1.4E-11). Further GSEA characterised this module as highly associated to immune response and INFγ-activation (pFWER<0.01) (Figure 5D). A constructed network based on the intramodule connectivity of module 1 and 2 was presented in Figure 5E. All modules were connected by hub-genes like CTSO and LACTB, which were part of the immune response related to expression module 2.
Rationale of the study
Here we present an integrative analysis based on data of the TCGA database and glioblastoma multiforme samples of the Department of Neurosurgery, Medical Center Freiburg. The purpose of this study was to identify potential molecular or genetic alterations in glioblastoma multiforme influencing the PD-L1 expression. These functional alterations could be used in future diagnostics and help to identify patients who may profit from a PD-L1/PD-1 inhibition.
Firstly, there were significant differences in the expression levels of PD-L1 regarding the different glioma WHO grades, expression subgroups and molecular subgroups (Figure 1B). Lower-grade glioma (LGG, WHO II+III) showed low expression level of PD-L1 (Figure 1A) compared to glioblastoma multiforme. Secondly, IDH1/2 wild-type subgroup (Figure 2A) showed similar PD-L1 expression as found in glioblastoma multiforme. This finding supports the recently published hypothesis that IDH1/2 wild-type subgroup represents typical glioblasoma multiforme-like genetic alterations .
Finally, after performing an analysis of frequently occurring genetic alterations in glioblastoma multiforme, NF1 loss-of-function was significantly associated with strong PD-L1 expression in both TCGA and Freiburg databases.
NF1 loss-of-function and RAS-MAPK pathway
The integrative analysis of glioblastoma multiforme proteome data confirmed the correlation between RAS-MAPK pathway and PD-L1 expression (Figure 3D, 3E). Meanwhile, NF1 loss-of-function (NF1 mutation and NF1-Deletion) resulted in an activation of MAPK pathway, which affected down-stream targets like MAPK, MEK and JNK (Supplementary Figure 1), thus suggesting a putative impact of NF1 on the PD-L1 expression. The strong association of MAPK pathway activation in patients with NF1 deletion was validated by immunostainings of the Freiburg cohort (Figure 4). Equally to the WPCNA, a strong correlation between PD-L1 protein-level and p-MAPK14 was shown. Those findings were in line with recently presented results  and supported by further analysis identifying a strong correlation of PD-L1 expression with mesenchymal subgroup genes (Expression module 1, Figure 3E). Additionally, the latter have been recently shown to correlate with NF1 mutation [21, 24]. As known from the literature, PD-L1 is regulated by the STAT3 transcription factor, which is also a transcriptional master regulator of the mesenchymal subgroup [20, 25]. Parsa et al., 2007 reported an association between PTEN loss and a up-regulation of PD-L1 expression . This association was not found in our analysis which is in line with results reported by Berghoff et al., 2014 . Environmental factors as hypoxia within the up-regulation of HIF1A take part in the mesenchymal signature . This expression subgroup was found to be highly connected to immune response and up-regulation of PD-L1 expression within the tumour . In summary, this study reported a strong coherence between the immune related genes included PD-L1 and the mesenchymal signature of glioblastoma multiforme.
PD-L1 and immune response
Another interesting finding was the correlation of PD-L1 expression with the immune response/INFγ activation. This results were in-line with recently published works, showing that pro-inflammatory substrates such as INFγ result in an increased PD-L1 gene expression [5, 27, 28]. GBM are tumours with a large heterogeneity of different cell clones. One major cell population in glioblastoma multiforme was characterized as “immune-cluster” by single-cell RNA sequencing . This immune phenotype was similar to other solid tumours (liver, kidney etc.) . In general, glioblastoma multiforme are tumours that implement a strong immune response within different parts of the tumours and potentially targetable by checkpoint-inhibitors.
PD-L1 and IDH1/2 mutation
Patients affected by an IDH1/2 mutation showed a hypermethylated PD-L1 promoter, which epigenetically silences the PD-L1 gene expression (Figure 2C, 2D). Of note, recent works showed that treatment with hypomethylating agents in patients with small-cell lung cancer resulted in an increased expression of PD-L1  underlining the effect of epigenetic regulation on PD-L1 expression. Epigenetic regulation of PD-L1 is also known for many other cancer types .
Limitation of the study
This study has also some limitations. Firstly, the small sample size and the disregarded heterogeneity could have led to false-positive associations or confounder effects. However, conservative statistical methods with corrections for multiple testing at each level of analysis were applied. Only family-wise error corrected values are reported for the sake of robustness in enrichment analysis.
MATERIALS AND METHODS
TCGA data platform
Public available Level 3 TCGA (https://tcga-data.nci.nih.gov/tcga/) data were used for analysis. Data were downloaded at the UCSC Cancer Genome Browser. Expression analysis was performed based on Agilent array data (TCGA GBM G4502A) for high-grade glioma and RNA-seq data (TCGA LGG HiSeqV2 PANCAN) for low-grade glioma. Both datasets were normalized and log2 transformed. For mutation analysis the PANCAN AWG gene-level mutation data set was used for further analysis. Proteomic data were taken from TCGA database as normalized level 3 RPPA data.
Tissue collection and histology
Tumor tissue was sampled from contrast enhancing regions identified by intraoperative neuronavigation (Cranial Map Neuronavigation Cart 2, Stryker, Freiburg, Germany) during tumor resection. The tissue was snap-frozen in liquid nitrogen immediately after resection and processed for further genetic/metabolic analysis. Tissue samples were fixed using 4% phosphate buffered formaldehyde and paraffin-embedded with standard procedures. H&E staining was performed on 4 μm paraffin sections using standard protocols. Immunohistochemistry was applied using an autostainer (Dako) after heat-induced epitope retrieval in citrate buffer. IDH1 mutation was assessed by immunohistochemistry using an anti-IDH1-R123 antibody (1:20, Dianova).
Genome wide expression data of 48 patients (Histology: Glioblastoma multiforme WHO Grade IV) (Department of Neurosurgery, Freiburg, Germany) was used for expression analysis in addition to TCGA data (Clinical data: Supplementary Table 1). RNA was prepared using the RNAeasy kit (Qiagen). An amount of 1.5 μg RNA was obtained for expression arrays analysis. Arrays were performed by human gene ST 2.0 chip (Affymetrix). Raw data were processed, normalized and controlled by R software and the Affymetrix R-package. Data were comparable to Level 3 TCGA data. Molecular subgroups were defined by machine-learning algorithm (random forest) based on TCGA classification (Supplementary Figure 2).
Genomic data processing
Whole-exome sequencing data (291 patients) of the TCGA database were processed as level III data. Co-mutation analysis was performed in R software and free-available implemented packages. Additional, copy-number variants from 577 patients (Level 3 TCGA data) were analysed. 45 Patients of the Freiburg validation cohort were analysed by Genome-Wide Human SNP Array 6.0 (Affymetrix) and pre-processed by the genome core facility in Freiburg (Department of Hematology, Oncology and Stem Cell Transplantation).
Weighted gene co-expression analysis
Expression analysis was performed on 585 patients (Array by AgilentG4502A). WGCNA uses the topological overlapping measurement to identify corresponding modules. These modules were analysed by their eigengene correlation to PD-L1 expression. The WGCNA analysis is a robust tool for integrative network analysis and was used in several recent studies [31–33]. WGCNA established networks were exported to Cytoscape 2.0  for further visualisation. The WGCNA integrated function (exportNetworkToCytoscape) was used to calculate a weighted network. A detailed description of WGCNA was recently reported .
Gene set enrichment analysis (GSEA)
Permutation based gene set enrichment analysis (GSEA) was performed for each module to find specifically enriched biological functions and related pathways . Pre-ranked GSEA were performed with 1000 permutations. P-values were calculated by familywise error rate (FWER), which is a robust method for multiples testing . The Molecular Signatures Database version 5.0 was used including pathways gene sets (C2) (http://www.broadinstitute.org/gsea) as input databases for this analysis. GSEA plots were visualised by limma R-package (barcodeplot function).
Weighted proteome correlation network analysis
Protein data of 215 patients were extracted from the TCGA database. WPNA was performed for proteome data of the TCGA database and PD-L1 expression. WPCNA was performed with the WCNA-package (R-software) as described above. Modules were built by topological overlapping measurement. The power for network analysis was adapted until scale-free topology was achieved. Derived modules were analysed by their eigengene correlation to PD-L1 expression level. Additional networks were created based on intermodule connectivity in Cytoscape 2.0 as described above.
Methylation analysis of PD-L1 promoter
Level 3 methylation data of the TCGA database (155 patients, Methylation 450k) was used for an epigenetic analysis of PD-L1 promoter. Analysis of the correlation coefficient (Pearson correlation, Fischer’s Exact test) between expression and methylation values of all CpG sides of the PD-L1 promoter was performed as described for MGMT promoter by Bady et al. 2016  (Figure 2C). CpG side “cg19724470” was found as significant negatively correlated to PD-L1 expression and used for further validation of PD-L1 promoter methylation (Figure 2D).
Overall survival (OS) was available for 45/48 patients (no event for 9 patients, censored). The Kaplan-Meier method was used to provide median point estimates and time-specific rates. The Hazard-Ratio (HR) was calculated by Cox-Regressions tests. Analysis was performed by survival package included in R-Software.
The following antibodies were used in immunostaining analyses: phospho-JNK (Phospho-SAPK/JNK (Thr183/Tyr185), Cell signaling, 1:200), p-MAPK14 (p-p38 Antikörper (Thr 180/Tyr 182), Santa Cruz 1:200), PD-L1 (E1LRN, Cell signaling 1:200) and phospho-Akt (Phospho-Akt (Thr 308), Cell signaling, 1:200). Primary antibodies were used at the concentration indicated by the manufacturers. Anti-Mouse and anti-Rabbit Alexa488- or Alexa594-conjugated (Life Technologies) were used as secondary antibodies. Alexa594-conjugated antibodies were used at 1:200 dilution and Alexa488-conjugated antibodies were used at 1:100 dilution. Pictures were acquired using a fluorescent microscope (FL10i, Olympus). Image quantification was performed by ImageJ and analyzed by R-software.
In conclusion, this study reported a comprehensive analysis of PD-L1 expression in glioblastoma multiforme. PD-L1 was highly associated with the mesenchymal subgroups and in line with mesenchymal-like genetic alterations like a loss-of-function in NF1. Patients with a NF1 mutations/deletion or RAS-MAPK pathway activation may profit from a targeted therapy with PD-1/PD-L1 inhibition.
The results published here are in part based upon data generated by the TCGA Research Network: http://cancergenome.nih.gov.
CONFLICTS OF INTEREST
The authors state no conflicts of interest.
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22. Brat DJ, Verhaak RG, Aldape KD, Yung WK, Salama SR, Cooper LA, Rheinbay E, Miller CR, Vitucci M, Morozova O, Robertson AG, Noushmehr H, Laird PW, et al. Comprehensive, integrative genomic analysis of diffuse lower-grade gliomas. N Engl J Med. 2015; 372:2481–98. doi: 10.1056/NEJMoa1402121.
23. Turcan S, Rohle D, Goenka A, Walsh LA, Fang F, Yilmaz E, Campos C, Fabius AW, Lu C, Ward PS, Thompson CB, Kaufman A, Guryanova O, et al. IDH1 mutation is sufficient to establish the glioma hypermethylator phenotype. Nature. 2012; 483:479–83. doi: 10.1038/nature10866.
24. Yoshihara K, Wang Q, Torres-Garcia W, Zheng S, Vegesna R, Kim H, Verhaak RG. The landscape and therapeutic relevance of cancer-associated transcript fusions. Oncogene. 2015; 34:4845–54. doi: 10.1038/onc.2014.406.
25. Carro MS, Lim WK, Alvarez MJ, Bollo RJ, Zhao X, Snyder EY, Sulman EP, Anne SL, Doetsch F, Colman H, Lasorella A, Aldape K, Califano A, et al. The transcriptional network for mesenchymal transformation of brain tumours. Nature. 2010; 463: 318–25. doi: 10.1038/nature08712.
26. Joseph JV, Conroy S, Pavlov K, Sontakke P, Tomar T, Eggens-Meijer E, Balasubramaniyan V, Wagemakers M, den Dunnen WF, Kruyt FA. Hypoxia enhances migration and invasion in glioblastoma by promoting a mesenchymal shift mediated by the HIF1α-ZEB1 axis. Cancer Lett. 2015; 359: 107–16. doi: 10.1016/j.canlet.2015.01.010.
27. Kondo A, Yamashita T, Tamura H, Zhao W, Tsuji T, Shimizu M, Shinya E, Takahashi H, Tamada K, Chen L, Dan K, Ogata K. Interferon-gamma and tumor necrosis factor-alpha induce an immunoinhibitory molecule, B7-H1, via nuclear factor-kappaB activation in blasts in myelodysplastic syndromes. Blood. 2010; 116:1124–31. doi: 10.1182/blood-2009-12-255125.
28. Sznol M, Chen L. Antagonist antibodies to PD-1 and B7-H1 (PD-L1) in the treatment of advanced human cancer. Clin Cancer Res. 2013; 19:1021–34. doi: 10.1158/1078-0432.CCR-12-2063.
29. Patel AP, Tirosh I, Trombetta JJ, Shalek AK, Gillespie SM, Wakimoto H, Cahill DP, Nahed BV, Curry WT, Martuza RL, Louis DN, Rozenblatt-Rosen O, Suvà ML, et al. Single-cell RNA-seq highlights intratumoral heterogeneity in primary glioblastoma. Science. 2014; 344:1396–401. doi: 10.1126/science.1254257.
30. Wrangle J, Wang W, Koch A, Easwaran H, Mohammad HP, Vendetti F, Vancriekinge W, Demeyer T, Du Z, Parsana P, Rodgers K, Yen RW, Zahnow CA, et al. Alterations of immune response of Non-Small Cell Lung Cancer with Azacytidine. Oncotarget. 2013; 4:2067–79. doi: 10.18632/oncotarget.1542.
31. Langfelder P, Horvath S. WGCNA: an R package for weighted correlation network analysis. BMC Bioinformatics. 2008; 9:559. doi: 10.1186/1471-2105-9-559.
32. Holtman IR, Raj DD, Miller JA, Schaafsma W, Yin Z, Brouwer N, Wes PD, Möller T, Orre M, Kamphuis W, Hol EM, Boddeke EW, Eggen BJ. Induction of a common microglia gene expression signature by aging and neurodegenerative conditions: a co-expression meta-analysis. Acta Neuropathol Commun. 2015; 3:31. doi: 10.1186/s40478-015-0203-5.
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35. Heiland DH, Mader I, Schlosser P, Pfeifer D, Carro MS, Lange T, Schwarzwald R, Vasilikos I, Urbach H, Weyerbrock A. Integrative network-based analysis of magnetic resonance spectroscopy and genome wide expression in glioblastoma multiforme. Sci Rep. 2016; 6:29052. doi: 10.1038/srep29052.
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All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 License.

domain: health


Sao Paulo, Nov 5: Fred scored his league-leading 17th goal of the season as Fluminense held Sao Paulo to a 1-1 home draw on Sunday and increased its lead over Atletico Mineiro in the Brazilian championship.
Fluminense moved closer to its second title in three years by opening a nine-point advantage with four rounds left. Ronaldinho's Atletico Mineiro had its chances significantly diminished after a 1-0 loss to Coritiba.
Palmeiras, the Brazilian team with the most national titles, is in serious danger of relegation after being held 2-2 at home by Botafogo. Argentina striker Hernan Barcos scored both goals for Palmeiras, while Uruguay midfielder Nicolas Lodeiro netted one of Botafogo's goals and set up the other.
Argentina forward Juan Martinez scored in Corinthians 2-0 win over Atletico Goianiense, which was demoted to the second division with the loss.
On Saturday, Neymar scored three goals and set up the other in Santos' 4-0 routing of Cruzeiro, receiving standing ovations even from Cruzeiro fans.
Fluminense added another point to its lead by containing fourth-place Sao Paulo at the Morumbi Stadium in Sao Paulo.
Luis Fabiano opened the scoring for Sao Paulo in the 50th after a mistake by Fluminense defender Gum, who mishit the ball while trying to pass it back to goalkeeper Diego Cavalieri. The striker got to the ball and easily dribbled past Cavalieri to find the open net.
Fred equalized when Sao Paulo defender Rafael Toloi erred in the 68th, allowing forward Samuel to steal the ball away from him and set up Fred for the easy goal from near the penalty spot. The striker has one goal more than Luis Fabiano.
Fluminense won despite playing without former Chelsea and Portugal playmaker Deco, who is nursing a right thigh muscle injury. Sao Paulo, which moved closer to securing a Copa Libertadores spot in 2013, had won five of its past six matches. The top four teams will play in the Copa Libertadores in 2013.
Third-place Gremio won 1-0 at home against Ponte Preta thanks to a last-minute goal at the Beira-Rio Stadium in Porto Alegre on Saturday. Striker Andre Lima scored the winner with a header from a corner taken by World Cup veteran playmaker Ze Roberto. Gremio, coached by Vanderlei Luxemburgo, is 10 points off the lead but also has virtually secured a Copa Libertadores berth.
On Saturday, Cruzeiro fans chanted Neymar's name and applauded the striker after he scored his hat trick in Santos' 4-0 win at the Independencia Stadium in Belo Horizonte. Neymar scored in the 11th, 35th and 83rd minutes and gave the pass for Felipe Anderson's goal in the 53rd.
“I'm moved after seeing this type of reaction from Cruzeiro fans today,” said Neymar, who has 12 goals in this year's Brazilian league. “I will keep this with me to the rest of my life. Today I can consider Cruzeiro my second home and I have to thank all the fans for their support.”
The result snapped Santos' three-match winless streak, but last year's Copa Libertadores champion is only ninth in the 20-team standings.
Eight-time national champion Palmeiras moved closer to relegation after the draw with Botafogo on Sunday. Barcos scored the equalizer in stoppage time, improving his goal tally to 13. He has a total 27 goals in 2012.
Uruguay midfielder Lodeiro scored Botafogo's first goal and set up the second. Coming off three straight wins, Botafogo was without 36-year-old Dutch midfielder Clarence Seedorf, who also is nursing a right thigh muscle. The team is fifth in the standings.
Palmeiras is 18th in the standings, seven points from safety with four rounds to go. It will face leader Fluminense at home in the next round.
Television images showed some disgruntled Palmeiras fans trying to invade the pitch after the match and police had to use rubber bullets to contain them. Local media said two police officers were slightly injured.
Palmeiras is still trying to annul its 2-1 loss to Internacional because the referee allegedly used TV images as the basis for his decision to disallow what could have been a second-half equalizer. A sports tribunal will hear the case on Thursday.
Atletico Goianiense became the first team relegated after the home loss to Corinthians, which is focused on the Club World Cup in December and is only eighth in the standings. Martinez scored Corinthians' first goal.
On Saturday, Flamengo moved farther from demotion with a 1-0 win over Figueirense in Volta Redonda. Hernane scored Flamengo's 73rd-minute winner. Figueirense, in
second-to-last place and virtually relegated, had two first first-half goals disallowed after controversial offside calls.
The bottom four clubs are be demoted. Bahia, the first team out of relegation zone, edged Portuguesa 1-0, while Sport—four points from safety—defeated seventh-place Vasco da Gama 3-0 at Sao Januario Stadium, handing the Rio de Janeiro club its sixth loss in a row. Vasco playmaker Juninho, who played for Brazil in the 2006 World Cup, was seen making obscene gestures to Sport fans during the match.


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This articulated dump truck is built to provide up to 205kW or 274 horse power, delivered from its powerful Mercedes-Benz engine to the 18.4t B 25 D machine. The B 25 D is categorised in the smallest machine segment in the articulated dump trucks category. The model's dimensions are 9.501m x 2.88m x 3.425m. Since 2001, this model has included Aircondition.
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Consult Bell Equipment Co SA's B25D Hooklift Truck brochure on DirectIndustry. Page: 1/2. SELL YOUR PRODUCTS Under our policy of continuous improvement, we reserve the right to change specifications and design without prior notice. Photographs featured in this brochure may include optional equipment. Articulated Dump Truck B50D. 6 Pages
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City is seeking alternate locations
The majority of the Driggs disc golf course will be temporarily defunct, the city announced in March, after the property owners decided to terminate the lease on the 8.5-acre lot that the city has been using as an extension of the 5th Street Park.
The disc golf course, designed by Karen and Jazz Russell and installed in 2003, has long been a haven for residents seeking convenient shoulder season recreation. The city leased the eastern 8.5 acres, which were once county-owned land and included a state gravel pit, essentially for free. In 2008 the property was auctioned off to Cottonwood Hollow, an LLC owned by Squeak Melehes and Deb and Dana Mackenzie, and the city continued its lease.
Melehes renegotiated the lease last spring and the city agreed to pay $125 per month so that the owners could recoup some of the property taxes they’ve paid on the vacant lot in the past decade.
Now Melehes plans to build a house on the southwest corner of the property, although he has expressed interest in donating two or three acres that comprise the old gravel pit, according to Driggs community development director Doug Self.
The city will be removing the disc golf baskets on the private property as soon as the snow melts and putting up signs discouraging trespassing on the short section of trail that winds through the trees at the back of the lot. The portions of the course that are on city property will remain for the time being, as other nearby options are explored.
The course could soon shift north. Ball Ventures, a real estate investment group in Idaho Falls, has stated it will donate the five acres of land on the corner of 5th and Little to the city as a future aquatic center site, and a nine-hole course could be established there as soon as early summer.
Self said there is also the possibility of a larger 18-hole course on the former Teton County landfill site, off Cemetery Road adjacent to the newly-established Teton Regional Land Trust conservation easement next to Teton Creek. A disc golf course on a reclaimed landfill is not unusual; for instance, the Freeman Park course in Idaho Falls is situated on an old garbage dump.
“The City recognizes the importance of the disc golf course to the community and will be working hard to transition to a new course as quickly as possible,” Self wrote in an email to the TVN. “The City also greatly appreciates the generous lease that Squeak Melehes and Dana and Deb MacKenzie provided for the course in the past.”


text:
One of the most hard working emcees straight out of Columbus, Ohio is none other than Dominique LaRue. Her latest project “Almost There” sees her teaming up with veteran producer J.Rawls who produces the entire project. On here, we see Dominique LaRue exploring different facets of what makes her tick. Almost There features a handful of guest appearances by Boog Brown, Apani b fly, Copywrite and more.
Tune in and get familiar right away.

domain: artistic


text:
We are a company based in Argentina, with a team working developing websites and graphic design pieces.
We are looking for sales agents ready to offer our quality design products, mainly graphic design, website design, professional photography and video.
We offer a good comission-based salary.
We need people with good organization and communication skills and willing to work in a independent growing project.
• Contact new clients and offer our services
• Be the personal contact with the client to get the requirements, updates and payments.
• Be proactive and manage multiple accounts to offer new services
We want to expand our services to different locations around the world.
Contact us for further information.
This is not a creative design job. It is a sales job.


text:
Gluten is what gives bread its elasticity and makes it rich and chewy. It is an indigestible protein found in grains that can cause the immune system to malfunction and attack the villi in the small intestine in people with celiac disease, causing major inflammation. For sufferers, gluten-free diets not only help ease their pain but can literally be life-saving.
Yet despite the fact that only 1% of people have celiac disease, sales of gluten-free products in Canada have more than quadrupled since 2008 and now top $800 million. Even Ryan Gosling has gotten into the act. One dietician estimated that 9 million Canadians are consuming gluten-free foods. But should they be?
Benefits of a Gluten-Free Diet for Non-Celiac Sufferers
If you don’t have celiac disease, you may still have what’s known as non-celiac sensitivity. Your symptoms wouldn’t be nearly as severe, of course, but they can be painful and inconvenient nonetheless. You might simply have an allergy to gluten, or, by coincidence, to other ingredients that tend to be in products that also contain gluten, such as wheat. There’s also the possibility that a person could be experiencing a placebo effect: if they’re convinced they have gluten sensitivity, not consuming it might legitimately make them feel better.
Also Helps with IBS
If you have Irritable Bowel Syndrome, you may also find benefits to not consuming gluten. While the connection between IBS and gluten is not fully understood yet, research has shown that a gluten-free diet can help reduce IBS symptoms, including diarrhea. It’s possible that IBS sufferers may have a sensitivity to gluten. It’s also possible that a gluten-free diet simply encourages those afflicted to stay away from foods that may cause irritation and pushes them towards more easily digestible choices.
A Gluten-Free Diet May Encourage Better Eating Habits
Since gluten can be found in grains that are used to make cakes, pastries, and other desserts, maintaining a gluten-free diet can help a person avoid detrimental foods. Fruits and vegetables, on the other hand, contain no gluten. People on gluten-free diets are therefore likely to consume less sugary foods and more healthy ones. In this way, avoiding gluten acts more like a guideline that gets people to watch what they eat and to avoid sugar – the real culprit.
Gluten-Free Mistakes You Should Never Make
Gluten-Free Doesn’t Automatically Mean Healthy – As noted above, gluten itself does not cause weight gain, though consuming too many carbs or too many sugary foods can. At times the substitutes for gluten can actually make a product less healthy than the original. Like with all foods, you have to check the label and shouldn’t assume that just because something is gluten-free that it’s healthy. Some gluten-free foods may in fact contain a lot of fat. Remember: excising gluten will not, on its own, make you lose weight.
You May be Missing Out on Important Nutrients – Whole grains can be very healthy, and if you’re not consuming them, you may have to supplement to make up for the benefits you’re losing out on. Gluten-free foods lack major nutrients, such as iron, calcium, riboflavin, phosphorous, and B12. A normal, healthy diet that includes grains, fruits, and vegetables, and avoids fatty and sugary foods, would probably be a superior option. Before you embark on a gluten-free diet, it may be best to consult with a doctor or nutritionist.

domain: food


Our local development databases do not have user avatar images. But that makes our forum pretty empty. So here is my avatar generator that will generate an avatar for a given id.
Started wanting to use a normal avatar type meme. But I can't draw, so defaulted to something others have drawn. Monsters.
There are a ton of great monster generators out there.
Here is where I got my start
- [http://www.splitbrain.org/projects/monsterid][Origional Monster Id]
- [http://scott.sherrillmix.com/blog/blogger/wp_monsterid-update-hand-drawn-monsters/][wp_monsterid] (Great images)
- [http://scott.sherrillmix.com/blog/blogger/wp_identicon/][wp_identicon] - identicons. nice work with color choices
- [http://blog.gravatar.com/2008/04/22/identicons-monsterids-and-wavatars-oh-my/][gravatar] - summary article
- [http://blog.dmcleish.id.au/2007/12/20/wavatar-artwork/][wavatar] - newer art work
- [http://scott.sherrillmix.com/blog/blogger/wp_monsterid-update-hand-drawn-monsters/][newer monsters] - newer art
Using the very pretty wp_monsterid/faces Not using their algorithm. so may need to tweak in the future


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My daughter just turned 11. She’s stretched so tall; she seems to be sprouting wings. She now stares at me each time I mispronounce a word from the storybook. I can see the dimming in her eyes. My mama isn’t perfect. My younger daughter asks me a question. She pricks her ears up. And I could no longer guess or hatch a plot. She’s listening for the truth. Hanging on to my word. “Does my mother truly know, or is she merely making things up?”
I lent her my phone. Deleted YouTube and Safari and yet permitted her Viber. Sometimes I see her messaging her friends. They label it SCOOPS. I humor myself. It’s literature and cute, and she’s learning letters. But I’ve been quite erratic about how much free rein or restraint I give. On some days, she’s rid of a mother bird looking over her shoulder. On other days, I wrestle with: have I granted her too much autonomy? Her wings are unfettered, and too soon.
How do you choreograph this brand new dance? And I’m afraid my books only supplied me enough wisdom till the 10th mark. She seems neither child nor adult. Not yet an adolescent. She’s a sapling. Slender and too young and yet quite ready for transplanting. She’s grown wings and if she stepped farther away from our nest, I know she could fly. Clumsily onto a branch, wing her way into the garden, but then flit back again.
She still plays make-believe. Today she invented a wizard and witch school. She’s wearing make-up. Her eyes are perfectly lined with kohl; her eyelids shadowed a misty blue. I don’t own lipstick. Where did that misty blue shadow come from? Who taught her how to paint herself like that?
I teach her math. She giggles at my mistakes. Shows me how to do long division.
Already she’s discovering worlds on her own. Already there are tricks that are were not mine to teach. Lessons where I was not the teacher.
She still believes in St. Nicholas. I wondered about that. Wondered whether she had a hint, or if she was merely bluffing for a trinket inside her stocking. And yet her eyes still sparkled as she reached down and pulled up a toy. I see her playing with it. Her dolls still talk. They have tea. She still fabricates them a bed and cucumber eye patches so they can wake up without eye bags. And when we paint, she still colours her trees with leaves of rosy peach and her sky is a sapphire blue streaked with orange.
This life with her is rare, and short, and miraculous. The clock ticks, our world spins, and it’s another turn around the sun.
I’m afraid next year my little bird will take wing higher and farther. Someday she’ll be hopping off to the piercing call of the bird next door. And her face painting will metamorphose to plumage feathers for a courtship ritual. Perhaps next year would be a year without Santa or talking dolls.
I don’t know what to do then. She will glide, spread her wings, and soar. Off to try trainer bras and even longer division. Off to learn about the world of dolled-up people who drink tea. Off to master the piercing call of the bird next door who might pierce her heart.
But I pray I’ve woven enough of magic and the sacred into her tapestry of life. That I’ve stitched and knotted and glued it tightly with wisdom, and compassion, and bravery. And that she remembers a nest that was carefully built with reverence, tied with laughter (and tears,) and cushioned with a whole lot of mud and feathers. And that wherever she soars, she will find that the trees are always praying with their rosy peach branches and that the sky remains a shimmering sapphire.
My daughter is 11 and she’s sprouting wings.


Rina Sawayama has dropped the visual for “Hold The Girl,” the title track of her upcoming sophomore album. In the video, directed by Ali Kurr, Sawayama she is seen in an old house, lamenting her personal pain, before climbing to a roof and jumping off. She lands safely in a bed, and is later joined by a crew, dancing in the house, before making a grand escape.
The song itself is a letter to a younger version of Sawayama, reminding her to give herself grace and care, as she grows and continues to make mistakes. “Hold The Girl” is one of the album’s many songs she wrote over the past year-and-a-half, dealing with the loneliness and heartache of the COVID-19 pandemic.
“‘Hold The Girl’ was the first song I wrote for the record at the end of 2020 — I had gone to therapy and had a revelation, so I decided to write this song… that was the start of it. I was crying before going into the studio to write about it,” said Sawayama in a statement accompanying the release of the single.
Check out the video for “Hold The Girl” above.
Hold The Girl is out 9/16 via Dirty Hit. Pre-save it here.


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As COVID-19 keeps people at home and away from their enterprises, the number of small business owners who are at risk of closing up shop is at unprecedented levels. Although this sector is an integral part of the U.S. economy, non-essential businesses have been forced to modify or cease operations in attempts to slow the global health crisis. The financial strain on small business owners, however, is a crisis in its own right.
“Sorry, We’re Closed.”
Long before the COVID-19 outbreak, small businesses were a substantial part of the country’s economy, generating 44% of U.S. economic activity.⁵ Today, their output is waning precipitously:
- More than 8 in 10 small businesses are concerned about the impact of COVID-19 on their business, with 58% being very concerned²
- 54% of all small businesses are closed or expect to close within the coming weeks²
- 43% of small business owners believe they have less than six months until a permanent shutdown is unavoidable,² and one in ten have less than a month
Government Programs Provide Temporary Relief
In response to the economic impact caused by the pandemic, the federal government passed the Coronavirus Aid, Relief, and Economic Security Act (CARES Act). Among its many provisions is a stimulus package designed for small business recovery. The Small Business Association (SBA) and their partnership with the nation’s lenders have been tasked with delivering critically needed financial assistance, including:
- Paycheck Protection Program – a loan to keep employees working that doesn’t have to be repaid if the workers remain on the payroll for at least eight weeks
- Economic Injury Disaster Loan – an advance of up to $10,000 to make up for a temporary loss of revenue that doesn’t need to be repaid
- Express Bridge Loan Pilot Program – loans of up to $25,000 to small businesses that have an existing relationship with an SBA Express Lender
- Debt relief – six months of forgiven principal, interest, and any associated fees that borrowers owe for all eligible loans
Delays are anticipated, so small business owners are expected to turn to lines of credit to cover costs in the interim. Yet a small business owner’s lack of credit knowledge and no established credit history can be a handicap to obtaining what they need:
- 42% of small businesses have no credit file or credit history⁶
- 72% of business owners don’t know their business credit scores⁶
- 42% of business owners who apply for financing are denied due to insufficient credit⁶
Additionally, even before COVID-19, small businesses were increasingly targeted by cyber criminals. They have lots of valuable data but lack the robust security infrastructure that larger companies possess to protect that data. In fact, the FBI recently issued a warning about the increase in cyber threats and fraud amid the COVID-19 pandemic7.
Small business owners are actively seeking assistance at this crucial time:
- 29% would like more guidance on how to keep their customers and employees safe²
- 26% want help on how to respond to the crisis²
Small Business Solutions from Experian Can Help
This is an opportunity for firms to provide their small business clients with business credit and identity protection solutions that can help them gain access to credit and protect against fraud during this critical time. Firms that do will see increased engagement with those clients and improved retention over the long-term.
With Experian’s Small Business Solutions, clients will have access to essential resources:
- Experian and Dun & Bradstreet credit file creation
- Business credit monitoring, identity protection, and fraud restoration assistance
- Business credit education center, so clients can learn about the unique factors that power their business credit score
Experian also offers identity theft alerts and dark web internet surveillance technology. Experian’s Business CyberAgent® scours websites, messaging services, blogs, bulletin boards, peer-to-peer sharing networks and IRC chat rooms to identify the illegal trading and selling of business information. Live customer service experts are ready to help small business owners manage and understand their credit concerns, as well as resolve any identity theft issues that arise.
Giving small business clients valuable assistance to protect their financial health in this time of crisis can not only create additional touch points with brands, but can be a saving grace. This generates long-term goodwill and, ultimately, client loyalty. As COVID-19 continues to wreak havoc, small businesses can use all the help they can get.
¹ NSBA, Small Business Poll, March 2020
²US Chamber of Commerce, Special Report on Coronavirus and Small Business, April 2020
³ CBA, Statement on SBA Small Business Stimulus Program
⁴WSJ, Big Banks Favor Certain Customers in $350 Billion Small-Business Loan Program, April 2020
⁵SBA, Small Businesses Generate 44 Percent of US Economic Activity, January 2019

domain: finance


text:
* ESC - закрити вікно пошуку
Vogue UA releases a book with the most iconic pictures of the last 10 years12.08.2022
The team of Ukrainian fashion magazine is to present a book “9 ½ years of Vogue in Ukraine”, which collects the most iconic photographs from the pages of Vogue UA since its launch. This has been announced on the magazine’s website.
This edition features iconic pictures from our archives that span a decade of Vogue UA storytelling. It includes portraits of Ukrainian and international celebrities, cultural elite, activists and athletes.
“In March 2022 we made a hard decision to temporarily suspend the magazine’s print edition and recalibrate our online platforms adjusting to the new reality of war. We believe that keeping our voice alive and loud is a way of resistance. One of the most ambitious anniversary projects, is a collectable book edition showcasing the best of what the Vogue UA team has created over the decade. So instead of folding this book project altogether, we, interrupted, chose to publish the anthology and to title it ‘9 1/2 Years of Vogue in Ukraine’. The missing half of the year is a reference to the tragic events in Ukraine that put our operations on hold, and to the present that has been stolen from us,” says Philipp Vlasov, Vogue UA Editor-in-Chief.
Part of the profits from the sale will be donated to Kyiv’s Ivan Honchar Museum which restores and preserves Ukrainian cultural heritage.
The book will be released in September. Preorders are open now by the link.
It shall be reminded that the magazine publisher Condé Nast, that issues popular magazines GQ, The New Yorker, Wired, Vanity Fair, Vogue and others, breaks franchise agreement and ceases publishing in russia.
This publication is sponsored by the Chytomo’s Patreon community
що більше читаєш – то ширші можливості

domain: library


Dr. Marty Becker severs ties with ABC
A regular guest on Good Morning America and several other television shows, Marty Becker, DVM, told dvm360 today that he is ending his longtime relationship with ABC. This comes after Becker says producers of 20/20 misled him regarding an interview he gave for the show’s Nov. 22 segment, “Is Your Veterinarian Being Honest with You?” which suggested that practitioners push unnecessary vaccinations and procedures to build their bottom line.
“I will no longer work with the ABC network effective Nov. 27,” Becker says in his statement. “I’ve enjoyed the 17 years we worked together and made many lifelong friends along the way. I deeply appreciate all of the opportunities I had on Good Morning America to promote the things I love: pets, pet owners and my beloved profession. However, for a number of reasons, I have made the decision that it’s time I move on.”
Earlier this week Becker was still trying to decide if he wanted to continue working with ABC. He said friends encouraged him not to jeopardize the relationship that gave him a platform to talk about veterinary medicine. However, today that relationship came to an end. Senior producers involved with Friday’s 20/20 broadcast did not immediately return phone calls seeking comment.
Becker says he will continue to represent the veterinary profession in mainstream media. He is a contributor to The Dr. Oz Show on CBS, and he says he has interviews lined up with National Public Radio and other news outlets in the wake of the 20/20 report. He also remains a spokesperson for VetStreet, a pet owner education and communication website, as well as an associate at two veterinary hospitals in north Idaho.
“Thank you to the veterinary community that has supported me in my mission to educate pet owners, and to the millions of loyal viewers, many of whom I hear from via mail, e-mail or chance encounters in public,” Becker’s statement reads. “I most recently had a woman come up to me in a hotel in Iceland and say, ‘Aren’t you Dr. Becker, the veterinarian on Good Morning America?
“You’ll still find me on network TV in 2014 and beyond,” Becker concludes. “As they say, ‘Stay tuned!’”


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BCSDN, as part of the activities of the Western Balkan CSOs for Global Development project, has published two country papers on Kosovo and Bosnia and Herzegovina, analyzing the state of play with the civil society and the environment in which it operates in both countries.
The country papers give background on the countries’ socio-economic situation and recent history, the EU integration process, the state of play with the Official Development Assistance and the state of development of the civil society. The aim is to provide information for assessing the potentials for civil society’s meaningful involvement in development policies in Kosovo and BiH, and in meeting the countries’ commitments related to the EU accession in the area of development and humanitarian aid. The papers were developed on the basis of the 2016 findings from the monitoring conducted by the Kosovar Civil Society Foundation (KCSF) in Kosovo and the Civil Society Promotion Center (CPCD) in BiH, within the Monitoring Matrix tool developed by BCSDN and its members and partners.
The full papers are available for download here:
This policy brief is part of the activities of the Western Balkan CSOs for Global Development project, supported by the Austrian Development Cooperation, SlovakAid and HORIZONT3000

domain: sociology


text:
Graduates find it difficult to secure a job out of uni
MORE than 10% of new bachelor degree graduates had not found a job within four months of completing their studies compared to 8.6% in 2012, a new survey by Graduate Careers Australia has found.
The annual survey showed 18.1% of new bachelor degree graduates had found part-time or casual work within four months of graduating.
GCA executive director Dr Noel Edge said while the transition of new graduates from higher education into the labour force remained slower than in the years prior to the global financial crisis, their long-term prospects were very strong.
Anecdotal evidence also found that graduates who had been placed in internships or found part-time work during their studies were more likely to find work after they graduated.
Did you get a job straight out of uni?
This poll ended on 20 December 2013.
Yes, I applied for jobs before I left uni
No, it took me a couple of months
I didn't go to uni
This is not a scientific poll. The results reflect only the opinions of those who chose to participate.

domain: news


text:
Last week I’ve went to Eindhoven with the rest of the CAMBADA team to RoboCup DutchOpen 2012, to participate in the Middle Size League.
Through the week we made several improvements and have been able to achieve the 3rd place, after competing with other physically stronger robots from Germany, Japan, Holland and Iran. I had a really good time in Eindhoven, especially because the organization was fabulous. Can’t find a single thing to complain about.
Now we are in preparations to RoboCup 2012 (Mexico).
If you want to know more about the CAMBADA team, this is our website.
Also, there is a website about the robotic projects currently made in University of Aveiro here.
More info on the DutchOpen can be found here.

domain: geography


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The actor, who is still recovering from the poor performance of her film, 3 and link-up stories with a married costar, Dhanush, is now looking to make it big in B-town
Kamal Haasan’s daughter, Shruti Haasan is yet to make her presence felt in Bollywood. The actor made her debut with Luck opposite Imran Khan, but didn’t get the kind of response she had hoped for. Her second film, Dil Toh Baccha Hai Ji, with Madhur Bhandarkar too didn’t do any good to her career. Since then, the actor has been busy making a name for herself in Telugu and Tamil films.
Even though the dishy babe, who also dabbles in singing, is now a big name down south, but it looks like she still harbours dreams of making it in B-town. The latest buzz is that Shruti has now relocated to Mumbai for good. After house hunting for quite some time, the actor has finally found a 2-BHK apartment in the posh lanes of Bandra.
So what made Shruti make the big move; it’s not like she’s being flooded with tons of Bollywood offers! Wethinks the debacle of 3 has made her rethink her strategies. What do you think?
Read more on Shruti Haasan:

domain: celebrity


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1500 songs in 400 motion pictures
Khadar, who composed a number of hit songs in Malayalam music, had a profession spanning 5 many years. During this time he wrote 1500 songs in about 400 movies, lots of which grew to become immortal. He wrote songs for composers KJ Yesudas, P Jayachandran, S Janaki and lots of extra. Apart from this, he labored with the largest and hit producers of the Malayalam movie business.
authorities engineer earlier than showing in movies
Born on 25 December 1948 in a small place like Poovachal close to Thiruvananthapuram, Khadar was a authorities engineer earlier than changing into part of movies. In 1973, he entered the Malayalam movie business. He remained lively within the movie business until 2011.
Popular lyricist of Malayalam movies Poovachal Khadar handed away. He was infected with Kovid-19 and was present process remedy on the Government Medical College. Khadar, 73, was placed on a ventilator after he developed pneumonia.


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Mar 21, 2006 (CIDRAP News) – The World Health Organization (WHO) today confirmed seven human cases of H5N1 avian influenza in Azerbaijan, making it the eighth country with officially recognized human illnesses.
A WHO reference laboratory in Britain confirmed the infection in samples from 7 of 11 patients tested, ranging in age from 10 to 21 years, the WHO said. Five of those patients, ranging from 16 to 21 years old, died of the illness.
The cases push the WHO's count of fatal cases over 100 for the first time. The agency now lists 184 confirmed cases with 103 deaths. Two cases reported in Egypt in the past few days have not yet been confirmed by a WHO reference lab.
The WHO said six of the cases occurred in the small settlement of Daikyand in the Salyan rayon in southeastern Azerbaijan, a former Soviet state on the west side of the Caspian Sea.
A 17-year-old girl was the first to succumb to the disease, on Feb 23, the WHO reported. Her death was followed by that of her 20-year-old female cousin on Mar 3 and the cousin's 16-year-old brother on Mar 10. A 17-year-old girl who was a close friend of the family died Mar 8. All four young people lived together or near each other, the WHO said.
"The additional two cases in Salyan involved a 10-year-old boy, who has recovered, and a 15-year-old girl, who is hospitalized in critical condition," the WHO said. The seventh case was in a 21-year-old woman from the western rayon of Tarter, who died Mar 9.
Two more patients, from Salyan and the adjacent area of Neftchela, have been hospitalized with pneumonia symptoms and are being tested, the agency said.
WHO and Azeri investigators have not yet determined how the patients were exposed to the virus. Interviews with family members have not revealed any history of direct exposure to dead or diseased poultry in several of the cases, the agency said.
However, there is some evidence that local residents might have collected carcasses of numerous swans, dead for some weeks, as a source of feathers, the WHO reported. Adolescent girls and young women usually have the job of defeathering birds, and a WHO team is investigating whether this could have been the source of infection in Daikyand, where most of the patients were girls between the ages of 15 and 20.
Azerbaijan is the third country outside East Asia, after Turkey and Iraq, with confirmed human cases of H5N1. The other countries with recognized cases are Cambodia, China, Indonesia, Vietnam, and Thailand. Vietnam has had the most cases by far—93, with 42 deaths—but has reported none in recent months.
The H5N1 virus first turned up in Azerbaijan in wild birds in February, with confirmation on Feb 10. The WHO reported Mar 10 that authorities were looking into a possible human cluster of 10 cases. On Mar 14, the agency said three patients who died had tested positive for an H5 virus and that confirmation of H5N1 was highly likely.
Also today, Egyptian authorities reported two more human H5N1 cases, in addition to the two reported in the past few days, according to an Agence France-Presse (AFP) report.
A 17-year-old boy named Mohammed Mahmud Abdul Ghani Ghabash, from Gharibiyah province north of Cairo, was hospitalized Mar 19, according to Health Minister Haten al-Ghabali, the story said. He worked on his father's poultry farm, where 3,000 birds died of flu over the weekend, AFP reported.
In addition, a 30-year-old woman named Fatima Mohammed Yussef, from Qaliubiya, north of Cairo, has been hospitalized with avian flu symptoms, the story said. It said she had continued working with her chickens despite a ban on the activity after the H5N1 virus struck Egypt in mid-February.
The story gave no information about testing of the two patients.
Two other Egyptians recently tested positive for the virus in a US Navy laboratory in Cairo, according to previous reports. A 30-year-old woman died of the illness Mar 17, and a 28-year-old man was hospitalized but improving as of yesterday.
Mar 21 WHO statement on Azeri cases

domain: medical


- Harvey Barnes scored from 30 yards as Barnsley drew 1-1 with QPR
- Famara Diedhiou and Aden Flint netted as Bristol City beat Bolton 2-0
- Preston went to Hull and returned with the three points after a 2-1
Barnsley thought they had beaten QPR thanks to a Harvey Barnes wonderstrike only for Luke Freeman to equalise for the visitors with just three minutes of action remaining to force a 1-1 draw.
Midfielder Barnes, on loan from Leicester, picked the ball up 30 yards from goal and curled a deft shot past goalkeeper Alex Smithies.
The Tykes thought they were home and dry, but Freeman had other ideas as he cut in from the left wing to power home with an 87th-minute goal.
Harvey Barnes of Barnsley celebrates his opening goal in the draw with QPR on Tuesday
Luke Freeman to equalise for the visitors with just three minutes of action remaining
Bristol City beat a 10-man Bolton 2-0 as Famara Diedhiou and Aden Flint netted either side of the break.
Bolton have now failed to win in 10 Championship games this season, and they were not helped when Filipe Morais was sent off for a second yellow card. Last night’s loss leaves them rooted to the bottom of the table with a meagre two points.
Preston meanwhile, went to Hull and returned with the three points after a 2-1 win thanks to Callum Robinson and Tom Barkhuizen goals.
Fulham also gained an away win, beating Nottingham Forest 3-1 as Aboubakar Kamara, Neeskens Kebano Stefan Johansen all scored.
Elsewhere, Millwall beat Reading 2-1 and Norwich triumphed 1-0 away at Middlesbrough.
Bolton Wanderers’ Gary Madine applauds the fans at the final whistle


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Zak’s book Back To Basics For The Republican Party is an important addition to any Conservative bookshelf. It’s not a big book, but it will put you in touch with some roots you’d be hard pressed to believe given today’s lying media.
Look whats up in my Home State:
Here is a taste:
(3) The department shall create and maintain a registry for owners of assault weapons
and large capacity magazines who qualify for registration under section 4 of this 2013 Act.
The department may adopt rules concerning the administration of the registry, including but
not limited to renewal and revocation procedures and storage requirements for assault
weapons and large capacity magazines.
(4) The department may conduct inspections of registered owners of assault weapons and
large capacity magazines to ensure compliance with the storage requirements of section 4
of this 2013 Act.
SECTION 6. This 2013 Act being necessary for the immediate preservation of the public
peace, health and safety, an emergency is declared to exist, and this 2013 Act takes effect
on its passage.
I just sent my local state senator (D) a rather blistering response to HB3200 and the tale of Blacks in America under Democrat leadership, I will let you know what kind of response I get,If I actually get one from the POS.
I think my local reps ears will be burning when he reads ( most likely a fag aide actually) my question about HB3200. Again, I will let you all know if I get a real response and not a canned ham.

domain: law


text: There’s a lot to be said for stability, especially when we’re talking about your money. Morris Bank has been here, right here in Middle Georgia, since 1954. And we’re not going anywhere. Every day, our roots are growing deeper and our ties to the community stronger. We’re invested in our towns.

domain: economics


text:
Future Music Festival for Melbourne is strictly 18+. ID is required. Thanks to our friends at Future Music Festival. Kiss has a limited amount of SINGLE TICKETS for the Melbourne gig on 8th March 2015. ...
February 18, 2015
*ONLY TWO WEEKS TO GO* WORLD ELECTRONIC MUSIC COMPETITION AUSTRALIA WEMC Australia follows a hugely successful European launch in 2014, which saw the prize winner embark on a world DJ tour through Las Vegas, Dubai ...
“BigSmit, 23 year old DJ from Essendon. Looking for that goosebump track every day, from melodic house to everything deep and bouncy. Let the journey begin!” Tune in this Friday at 6 pm . To ...
January 21, 2015
This week Cook & Big Dip from StrictlyOZ HipHop have special guest Live in the studio “FRAKSHA” from Broken Tooth Entertainment Smash Bros Crew to chat about his new release Its Just Bars 2 and ...
November 28, 2014
Possibly the most ridiculous, amazing edition of 3Fold Radio EVAARRRRRRR!!!!! Super special guests D-Nox & Beckers, Boris Brejcha and Victor Ruiz will be on mixing duties for the inaugural Saturday night edition of 3Fold Radio. ...
September 16, 2014
Justin Faust has quickly emerged at the vanguard of Europe’s nu-disco-electro-dance sound which has taken over dancefloors and DJ charts during the past half-decade. His new album drops 19th September on Discotexas with the hit single ...
January 29, 2014
A staple name in big room high energy electronic music for over a decade, pioneer of the UK’s ‘Nu-Skool Breaks’ movement, and co producer of leftfield proto-dubstep hit ‘Creator’ by Santigold, FreQ Nasty is switching ...
November 20, 2013
INSATIABLE !!! INSATIABLE released through Universal Music Group and Famous Record Corporation in USA. INSATIABLE is on iTUNES, YOUTUBE and other links !!! Go to RhettMay.com.au that’s RhettMay.com.au for LOTS MORE INFO!!!! Or contact firstname.lastname@example.org – we ...


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The Race to Cure SarcomaTM Philadelphia is just around the corner! On October 30, 2016, patients, survivors, family, friends, healthcare providers and other concerned individuals will come together to raise awareness and funds for sarcoma research.
One of our volunteers and team captains, Emilie Joly, shares why she’s involved.
I care about supporting the Sarcoma Foundation of America and participating in their Race to Cure SarcomaTM Philadelphia because I lost my mom to leiomyosarcoma in 2013, just THREE months after her diagnosis.
When she was diagnosed, I scrambled to find out more information and treatment options. Before her diagnosis I had never heard of it. I struggled to find much information about leiomyosarcoma, and it seemed even more difficult to find treatment options in the Lehigh Valley/Philadelphia area.
My whole family, including my mom, felt pretty hopeless with the lack of information available. It’s important to me to help raise money for research in her memory because while it’s considered “rare,” millions of people are affected by sarcoma – patients, friends, family, communities and loved ones – and no one should feel as if their situation is hopeless. Please help me, and others affected by sarcoma, keep hope alive!
Together we will move mountains to make this happen.
Start a Team, Join an Existing Team or Race as an Individual.
Take the first step towards a cure for Sarcoma at the Race to Cure SarcomaTM Philadelphia, October 30, 2016, at Warminster Community Park.
Funds raised will help support sarcoma cancer research locally, at the Sarcoma Center at Fox Chase Cancer Center, and across the country.
You can still help take a step toward the cure by making a donation to the Race to Cure SarcomaTM Philadelphia.


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Guest Artist Concert: Heather Killmeyer at AU Music Department March 19
Monday March 19, 2018: Recital in 125 Goodwin Hall, 7:30pm
Heather Killmeyer enjoys a varied career as a recitalist, chamber and orchestral musician, and educator. She currently serves as Assistant Professor of Double Reeds at East Tennessee State University. She previously was on the faculty at the University of the Incarnate Word and maintained a large private studio in San Antonio, Texas.
Heather Killmeyer earned her Doctor of Musical Arts from the University of Southern California, where she studied oboe with Allan Vogel and chamber music with David Weiss and Yehuda Gilad. She completed her Master of Music with Stephen Caplan at the University of Nevada Las Vegas. She earned her Bachelor of Music from the Cincinnati College-Conservatory of Music as a student of Mark Ostoich, and was a recipient of the Marcel Dandois Award. She has had additional studies with Jared Hauser and Brenda Schuman-Post on oboe and Sharon Kuster on bassoon.
Last Updated: March 21, 2018


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Jeremih x Ty Dolla Sign The Light
Jeremih & Ty dolla sign releases the official video for the track ‘The Light’. Jeremih and Ty Dolla Sign’s upcoming joint album MihTy is certainly on our list of the most anticipated projects of this year.
Chris Brown, French Montana and Lil Wayne are set to appear on MihTy which will have a little more than 10 songs, all produced by Hitmaka aka Yung Berg.
Download Jeremih x Ty Dolla Sign The Light Video;
Latest posts by Eric.O (see all)
- [MUSIC] Teni – Sugar Mummy - April 18, 2019
- SZA – Power is Power Ft. The Weeknd & Travis Scott - April 18, 2019
- [MUSIC] Olamide – Spirit - April 18, 2019


So far, Duke fans are feeling great about the season.
In Atlanta, Duke knocked off defending champ Kentucky. After that, the Blue Devils bounced Minnesota and VCU from the Bahamas to make it to the championship game of the Battle 4 Atlantis Tournament. There, Duke defeated second-ranked Louisville in a hard-fought game where every starter scored in double digits.
Early concerns about Seth Curry’s injury, the point guard play, and whether or not Mason Plumlee could live up to his physical promise all seem to have been answered in the affirmative.
With sports, however, there is always another obstacle in one’s path. And in Duke’s case, the impending clash with Ohio State in the ACC-Big Ten Challenge may be the toughest test yet.
The Blue Devils, brimming with confidence, are looking to defend home court against a nonconference opponent for the 97th straight game and at the same time avenge last season’s blowout loss in Columbus.
In order to beat Ohio State in the ACC-Big Ten Challenge, Duke must make sure these five things occur.


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Crochet the Small Miniature Pumpkins
This small miniature pumpkin is part of a Halloween Wreath by Yarnspirations. If following the pattern that I have provided for you, it’s the small pumpkin of the two that are in the pattern.
Judging by the size of my hands, you can see how tiny this is. It’s too cute!
Learn how to make this pumpkin with my free tutorial.

domain: design


Main / Medical / Dj chipman stickin n rollin
Dj chipman stickin n rollin
Name: Dj chipman stickin n rollin
File size: 242mb
22 Jan Stream Dj Chipman Stick It and Roll It by dj slider from desktop or your mobile device. 31 Mar Stream Frunt Back (Stickin' & Rollin') by DJ Chipman from desktop or your mobile device. Follow DJ Chipman and others on SoundCloud. Watch the video, get the download or listen to DJ Chipman – Stickin N Rollin for free. the full song is on youtube or any you can find baltimore club music.
Listen online to DJ Chipman - Stickin N Rollin and find out more about its history, critical reception, and meaning. Phone, Suggest a phone number Dj Chipman Stickin N Rollin. 2 likes. Music. Dj Chipman Stickin N Rollin. Posts about Dj Chipman Stickin N Rollin. 29 May Stickin n Rollin - Dj Chipman Download - This dat funk *You can copy and share with all your friends. Embed Code: Posted On: 29 May
Dj chipman stickin n rollin download. Click here to get file. Download woo tang song! Stickin n rollin dj chipman mp3 download. Jacksonville whoop woo get it!. Lyrics for Frunt Back (Stickin' & Rollin') by DJ Chipman. LyricsFrunt Back ( Stickin' & DJ Chipman Be the first to add the lyrics and earn points. Add lyrics . Check out Frunt Back (Stickin' & Rollin') by Dj Chipman on Amazon Music. Stream ad-free or purchase CD's and MP3s now on mosaicbarcelona92.com (Stickin N Rollin - DJ Chipman) Underground Dj going by the name DJ Chipman with hit singles like Peanut Butter Jelly Time, Wu Tang. Read more on mosaicbarcelona92.com Listen to all tracks and music by DJ Chipman for free. +. DJ Chipman Peanut Butter and Jelly Time free listening DJ Chipman Stickin N Rollin free listening.


text:
'Chriss - Cliffs' - Close to creek & beaches
Entire home hosted by Dallas & Cassie
- 6 guests
- 3 bedrooms
- 4 beds
- 2 bathrooms
Check yourself in with the lockbox.
90% of recent guests gave the location a 5-star rating.
Great check-in experience
95% of recent guests gave the check-in process a 5-star rating.
Every booking includes free protection from Host cancellations, listing inaccuracies, and other issues like trouble checking in.
Where you'll sleep
1 of 2 pages
What this place offers
Free parking on premises
Patio or balcony
Garden – Not fully fenced
Children’s books and toys for ages 0–2 years old, 2–5 years old and 5–10 years old
Unavailable: Carbon monoxide alarm
Select check-in date
Add your travel dates for exact pricing
4.87 out of 5 stars from 63 reviews
Where you’ll be
Hat Head, New South Wales, Australia
- 63 Reviews
- Identity verified
We are a family of 4 that love the beach and nature. Hat Head is our special place to relax and unwind.
During your stay
We are happy to help with tips on the area and as we dont live close by, we have our lovely neighbour who can assist you. This home is our special place and we love sharing it with guests who respect our simple house and come to explore and enjoy the local area. The location is in a quiet residential area and in respect of our neighbours we stress that strictly NO parties or noise at night will be tolerated. If you are planning a celebration with more than the max 6 guests we ask that you consider another property.
We are happy to help with tips on the area and as we dont live close by, we have our lovely neighbour who can assist you. This home is our special place and we love sharing it wit…
- Policy number: PID-STRA-5164
- Response rate: 100%
- Response time: within an hour
To protect your payment, never transfer money or communicate outside of the Airbnb website or app.
Things to know
Check-in: After 14:00
Check out: 10:00
Self check-in with lockbox
No parties or events
Health & safety
Airbnb's COVID-19 safety practices apply
No carbon monoxide alarm
Nearby lake, river, other body of water

domain: travel


text:
Amenities & Services
CLEAN, BUDGET INN
Whether you’re traveling for business or pleasure, it’s always nice to have a few of the comforts of home. And maybe even a few comforts you don’t have at home. The Sea Rock Inn has many amenities to make your stay relaxing and enjoyable.


Local celebrity Gil Rivera, Diamond Bar real estate agent.
Antiojitos Carmen Restaurant, Boyle Heights. Outtake for L.A. Mag.
Fresh Churros. Outtake from an LA Mag assignment.
Here’s a preview for a new project with a new friend, Rishi Kumar, from my hometown of Diamond Bar. He’s a sustainable suburban farmer/educator who’s turned every usable foot of yard-space at his suburban home into food growing space, where they produce food year round. Rishi has also been working with the nearby Pomona Unified School District where he’s helping to build gardens at some of the schools and always working on grant proposals for more.
Here’s a few photos from his home and two garden work days at the local elementary schools.
Top of the Mountain, Aspen 2013
In my holiday boredom I’m finally getting around to finalizing a bunch of scans from this past year. Here’s a fairly recent one from Aspen. Although I’ve never skied or snowboarded once in my life, when I was in Aspen for a couple days I bought a lift ticket so I could go up to the top, take some pictures, and come back down. This picture was taken just outside of the restaurant up there, The Sundeck, around 10 or 11 in the morning.
Daaaaang. So nice!
This is a job from USC/Dornsife College Mag that just came out. I was hired to photograph various aspects of the city of Pacoima in the San Fernando Valley for a story USC was doing on an environmental justice program (PERE) in the Dornsife College at USC.
Pacoima was listed as a “Toxic hotspot” community, where air pollution levels regularly exceed safety standards recommended by the state, along with five other lower income, mostly Hispanic communities. Maria Guzman (portrait) volunteered to monitor air pollution levels in Pacoima for PERE. The city is bordered on three sides by the 210, 5 and 118 freeways, while countless industrial businesses, auto shops and recycling plants line the streets. My job was to find the line where the urban and suburban meet and are forced to coexist.
One of the best assignments of the few I’ve had so far. It was also amazing that they requested I shoot film for the project, after seeing my series on my hometown. I spent about 5 days cruising Pacoima morning - afternoon. DREAM JOB