text:
Human Cancer Biology
Identification and Prognostic Significance of an Epithelial-Mesenchymal Transition Expression Profile in Human Bladder Tumors
Egbert Baumgart, Michael S. Cohen, Brasil Silva Neto, Micah A. Jacobs, Chad Wotkowicz, Kimberly M. Rieger-Christ, Andreia Biolo, Ron Zeheb, Massimo Loda, John A. Libertino and Ian C. Summerhayes
Egbert Baumgart
Michael S. Cohen
Brasil Silva Neto
Micah A. Jacobs
Chad Wotkowicz
Kimberly M. Rieger-Christ
Andreia Biolo
Ron Zeheb
Massimo Loda
John A. Libertino
Ian C. Summerhayes
DOI: 10.1158/1078-0432.CCR-06-2330 Published March 2007
Purpose: Epithelial to mesenchymal transition (EMT) is reportedly an important transition in cancer progression in which the underlying cellular changes have been identified mainly using in vitro models. In this study, we examined the expression pattern of EMT markers in vivo and determined the occurrence and clinical significance of these events in a series of bladder carcinomas.
Experimental Design: Eight hundred and twenty-five tumor samples from 572 bladder cancer patients were assembled in 10 tissue microarrays. Paraffin sections from each tissue microarray were subjected to antigen retrieval and processed by immunohistochemistry for the expression of E-cadherin, plakoglobin, β-catenin, N-cadherin, and vimentin.
Results: Pathologic expression of E-cadherin, β-catenin, plakoglobin, and vimentin were associated with the clinicopathologic variables of grade and stage with only the cytoplasmic localization of plakoglobin found associated with lymph node status. Associations between the aforementioned markers were found significant as determined by the Spearman correlation coefficient with N-cadherin showing no associations in this analysis. In univariate survival analysis involving patients who underwent cystectomy, the reduction or loss of plakoglobin significantly influenced overall survival (P = 0.02) in which the median time to death was 2 years compared with 4 years when a normal level of plakoglobin was recorded. When the analysis was done for cancer-specific survival, low levels of both plakoglobin (P = 0.02) and β-catenin (P = 0.02) significantly influenced survival.
Conclusion: The putative markers of EMT defined within a panel of bladder carcinoma cell lines were recorded in vivo, frequently associated with tumors of high grade and stage. Although multivariate analysis showed no significant influence of the EMT biomarkers on survival, alterations associated with plakoglobin were identified as significant prognostic features in these tumors.
E- and N-cadherin
plakoglobin
Epithelial to mesenchymal transition (EMT) is a process that was first observed in embryonic development (1–3) and has more recently been implicated as an underlying event in neoplastic progression (4–7). To date, the definition and occurrence of EMT in in vivo tumorigenesis remains controversial (6, 8); however, the conceptual framework embracing loss of epithelial markers and gain of mesenchymal markers has been reportedly associated with numerous cancers (9–11), often identified in cell lines established from tumors representing different grades and stage (10, 12). In this way, the discontinuous progression model observed within a panel of cell lines of common tissue origin may be closely linked to the differentiation status of cells. In most cases, the projected transition between epithelial and mesenchymal phenotypes is accompanied by increased motility and invasive potential.
With the advent of the establishment of in vitro models of EMT, elicited by the action of different factors on alternative cell types (13–18), the existence of EMT in progression becomes more credible. In addition, such models enable us to confirm events linked to EMT identified in discontinuous progression models and to discover further molecular events underlying this transition (7, 19). Alterations associated with the cadherin/catenin complex often feature at the center of EMT related to increased migration and invasion of cells (19). Such events will follow different courses in different models dependent on the expression profile of the cadherins in the targeted cell. In a discontinuous model of EMT in bladder cancer, we have identified loss or reduced expression of E-cadherin accompanied by the silencing of plakoglobin expression as late-stage events in bladder tumorigenesis (20). This change was linked to increased motility and invasion potential. In bladder carcinoma cell lines, loss of E-cadherin expression is accompanied by novel expression of N-cadherin, representing cadherin switching from an epithelial-specific classic cadherin to a mesenchymal-specific cadherin (21). Such EMT-related events accompany increased motility and invasion in bladder carcinoma cells. In a previous report, we have shown novel expression of N-cadherin in human bladder tumor tissue (22). In this study, using a tissue microarray format, we evaluated five biomarkers linked to EMT to determine the occurrence of expression of these molecules in vivo and the potential prognostic value of such events within a well-defined series of bladder carcinomas.
Cell culture. Human bladder cell lines RT4, RT112, HU456, BC16.1, CUBIII, 5637, PSI, HT1197, HT1376, SCaBER, EJ, KK47, J82, UM-UC-3, and TCCSUP were maintained in DMEM supplemented with 7.5% fetal bovine serum and penicillin/streptomycin.
Invasion assays.In vitro invasion assays were carried out using modified Boyden chambers consisting of Transwell (8-μm pore size; Corning Costar Corp., Cambridge, MA) membrane filter inserts in 24-well tissue culture plates. For invasion assays, the upper surfaces of the membranes were coated with Matrigel (Collaborative Biomedical Products, Bedford, MA) and placed into 24-well tissue culture plates containing 600 μL of NIH/3T3 conditioned medium (experimental) or plain DMEM (control). Cells (1 × 105) were added to each Transwell chamber and allowed to invade toward the underside of the membrane for 24 h at 37°C. Cells that pass through the membrane were fixed in methanol, stained with crystal violet, and counted under a light microscope.
Immunocytochemical staining of bladder carcinoma cell lines. Cells were grown on glass slides, washed with PBS, and fixed in 3.7% formaldehyde for 15 min at room temperature. Cells were then rinsed in three changes of PBS and permeabilized in 0.5% Triton X-100 in PBS. Following three washes in PBS, cells were stained for EMT biomarkers using the antibodies listed below. Immunostaining was done on an automated stainer (Autostainer Plus, DAKO Corporation, Carpinteria, CA).
Western blot analysis. Subconfluent dishes of cells were washed in PBS followed by lysis in hot sample buffer [2× ESB-0.08 mol/L Tris (pH 6.8); 0.07 mol/L SDS, 10% glycerol, 0.001% bromophenol blue, and 1 mmol/L CaCl2] and sheared through a 26-gauge needle. Lysates were then assayed for protein concentration using the bovine serum albumin method (Pierce, Rockford, IL). After determination of protein content, β-mercaptoethanol (1%) was added to each sample. Samples were boiled for 5 min, and protein was loaded in each lane of a 7.5% or 12.5% polyacrylamide gel. Proteins were transferred overnight onto nitrocellulose. Membranes were blocked in 10% milk in TBS with 0.05% Tween and incubated with primary antibody overnight at 4°C. Blots were washed in TBS with 0.05% Tween, thrice for 15 min each, and incubated with secondary antibody linked to horseradish peroxidase for 60 min at room temperature. Blots were then washed as described above and developed with an enhanced chemiluminescence kit (Amersham, Arlington Heights, IL).
Clinical samples. We have used tumor tissue samples from patients diagnosed with bladder cancer at the Lahey Clinic Medical Center between 1990 and 2005 under an institutional review board–approved protocol. Formalin-fixed, paraffin-embedded tumor tissues from patients were retrieved from the archives of the Pathology Department. The tumor stage was determined using tumor-node-metastasis classification and graded according to WHO guidelines.
Construction and immunohistochemical staining of tissue microarrays. Eight hundred and twenty-five tumor samples from 572 patients were assembled in 10 tissue microarrays. All tumor samples were transitional cell carcinomas. Tissue microarrays were designed with replicas for each tumor sample and each control. Controls included normal tissue from prostate, testis, tonsil, liver, cerebellum, kidney, lung, and bladder. Strategic placement of control cores in each tissue microarray enabled definitive orientation during the scoring of the tissue microarrays. Each tissue microarray consisted of 400 tissue cores (four cores per specimen) and control tissue cores for immunohistochemical validation and orientation. Multiple 4-μm sections were cut and stored at 4°C in the presence of a desiccant before immunohistochemical staining. Individual tissue microarray sections were deparaffinized and antigen retrieval in citrate buffer (pH 6.0; DAKO) was done. The antibodies were diluted with DAKO antibody diluent solution. Immunohistochemical staining was done on an automated stainer (Autostainer Plus, DAKO) using a high-sensitivity, polymer-based detection system (EnVision, DAKO). A negative control was included using a nonspecific mouse antibody solution (DAKO) substituting for the primary antibody.
Antibodies. Mouse monoclonal antibodies to cytokeratin, E-cadherin (DAKO), β-catenin, and plakoglobin (BD Transduction Laboratories, San Diego, CA) were diluted 1:25; 1:100, and 1:100 (working concentrations, 0.75, 7.56, and 2.5 μg/mL), respectively, for use in immunohistochemistry. Mouse monoclonal antibodies to mesenchymal cell markers N-cadherin (Zymed, San Francisco, CA) and vimentin (DAKO) were diluted 1:100 (working concentration 5 μg/mL) and 1:50 (working concentration 4.6 μg/mL), respectively. The negative control reagent (DAKO) is a cocktail of nonimmune mouse immunoglobulins (IgG and IgM) and was purchased prediluted. The specificity of each antibody in immunohistochemistry was determined using xenograft sections derived from bladder cell lines in which the expression profile of the antigen of interest had been previously identified.
Scoring of immunohistochemistry. The tissue sections were scored semiquantitatively, assessing staining intensity and protein localization, including membrane, cytoplasmic, or nuclear localization. In the case of E-cadherin, β-catenin, and plakoglobin, a staining intensity scale of 0 to 3 was applied. In normal bladder tissue cores, the staining intensity with each antibody was recorded as 3 with membrane localization throughout all epithelial layers of the mucosa. In tissue sections that displayed heterogeneous staining throughout the section or between cores from the same tumor, the worst-case-scenario score was assigned to that sample when >5% of the tumor cells displayed this phenotype. Greater than 90% concordance between scores from different cores of the same tumor was recorded. In samples where scoring differed between cores, the worst-case-scenario score was recorded for analysis. Specimens that exhibited a complete absence of staining or faint staining in <5% cells were scored negative. In the case of N-cadherin and vimentin, a positive or negative score was given for each tissue sample because the presence of either mesenchymal marker in bladder carcinoma cells represented novel expression. Each tissue microarray was scored independently (I.C.S. and M.L.). Where discordant results were obtained, both individuals rereviewed the stained cores to obtain a consensus.
Statistical analysis. Comparisons of groups were done by Pearson's χ2 test. Correlation between markers was determined by Spearman's correlation coefficient. Univariate overall and cancer-specific survival analysis was done using the product-limit procedure (Kaplan-Meier method), with the surgery date as the entry date. The log-rank (Cox-Mantel) test was used to compare survival curves for different categories of each variable. Variables with effect on survival in univariate analyses (P ≤ 0.15) were examined by log-log plot to determine how these variables could be incorporated into a Cox proportional hazards regression models, and variables with P ≤ 0.1 were maintained in the model. Test for interactions were carried out for the variables that were significantly related to survival in Cox regression analysis. Data were analyzed using the SPSS software package, version 11.5. B. Silva Neto and A. Biolo did the statistical analysis.
Characterization of EMT markers in bladder carcinoma cells. To establish the occurrence of the putative EMT phenotype in bladder carcinoma cells, we initially screened a panel of 15 cell lines for the expression of the five biomarkers we propose to evaluate in tissue sections. Table 1 shows the expression profile of each component as determined in Western blot analysis and includes morphologic and in vitro invasion data for each cell line. No nuclear localization of proteins was detected in cell lines using immunocytochemistry (data not shown). Morphologic classification of epithelial morphology was defined by tightly adherent cuboidal cells growing as discrete colonies with mesenchymal morphology defined by poorly adherent carcinoma cells displaying a stellate morphology. From the aforementioned data, it is clear that the projected EMT phenotype is recorded within this panel of bladder carcinoma cell lines and is correlated with a mesenchymal morphology and invasive phenotype as assessed in an in vitro assay.
Expression of EMT phenotype in bladder carcinoma cell lines
EMT phenotype in bladder tumor tissue. If the EMT phenotype identified in the bladder carcinoma cell panel is a valid paradigm in bladder cancer, we would expect to see the proposed modulations in epithelial and mesenchymal cell markers associated with more aggressive bladder disease. To test this postulate, E-cadherin, plakoglobin, β-catenin, N-cadherin, and vimentin expression levels and localization were assessed in tissue microarrays of primary bladder tumors. The construction of 10 microarrays included 825 transitional cell carcinoma samples from 572 patients. The tumor-node-metastasis and histologic grade of the samples is shown in Table 2 . Analysis by number of patients as opposed to number of samples revealed no difference in the results. In this study, the analysis of the superficial tumor group included pTa, pTis, and pT1 with the invasive group represented by pT2-T4 tumors. When T1G3 was included in the invasive group, the results did not change (data not shown).
Histologic and clinical variables of arrayed transitional cell carcinomas of the bladder
E-cadherin expression. A reduction or loss of E-cadherin expression was significantly correlated with pathologic tumor stage (P < 0.001), histologic grade (P < 0.001), but not lymph node involvement or lymphovascular invasion in bladder carcinomas. Loss or reduction in E-cadherin expression was recorded in 37.7% of superficial tumors and 60.2% of invasive tumors with similar values recorded for cytoplasmic localization of E-cadherin, 26% and 58.1%, respectively (Table 3 ).
Associations between EMT markers and clinicopathologic factors in patients with bladder cancer
β-Catenin expression. A reduction or loss of expression, in addition to a cytoplasmic localization of β-catenin, showed significant correlation with tumor grade and stage (P < 0.001) in bladder carcinomas. Neither the intensity nor the localization was found associated with a positive lymph node status or vascular invasion (Table 3). A reduction or loss of β-catenin expression was recorded in 16.7% of superficial tumors and 47.6% of invasive tumors (Table 3). The cytoplasmic localization of β-catenin revealed a greater differential between superficial and invasive tumor groups, 17.9% and 58%, respectively. Nuclear localization of β-catenin was recorded in 27 of 709 (3.8%) patient tumors, including 1 Ta, 9 T1, and 17 T2-T4 bladder tumors.
Plakoglobin expression. A reduction or loss of expression of plakoglobin and the cytoplasmic localization of plakoglobin was significantly associated with tumor grade and stage (P < 0.001). In addition, plakoglobin localization was also correlated with nodal status in bladder cancer patients (P = 0.04). Neither the level of plakoglobin expression nor the location was associated with vascular invasion. Loss or reduced plakoglobin expression was recorded in 15.2% of superficial tumors and 57.9% of invasive tumors. As with β-catenin localization, there existed a marked differential between the frequency of cytoplasmic localization of plakoglobin in the superficial and invasive tumor groups, 23.1% and 72.1%, respectively (Table 3). Nuclear localization of plakoglobin was recorded in 13 of 726 (1.8%) bladder carcinomas, including four Ta, four T1, and five T2-T4 lesions.
N-cadherin expression. Novel expression of N-cadherin was recorded in 61 of 746 (8.2%) bladder carcinomas (Fig. 1A and B ), including 32 superficial and 24 invasive tumors with 5 tumors that remained unstaged. Membranous N-cadherin staining was rarely recorded expressed throughout a tumor (Fig. 1B) but displayed a limited focal localization (Fig. 1A). Of the 61 tumor samples positive for N-cadherin expression, only 21 were scored as positive in all scorable tumor cores from a single patient. Novel expression of N-cadherin did not show a significant association with grade, stage, lymph node involvement, or vascular invasion.
Immunohistochemical staining showing N-cadherin (A and B) and vimentin (C and D) in low (A and C) and high-grade (B and D) bladder tumors. A, novel expression of N-cadherin in a superficial tumor showing focal localization within the tumor. B, N-cadherin expression throughout an invasive bladder tumor. C, absence of vimentin expression in epithelial cells of a superficial bladder tumor. D, novel vimentin expression in carcinoma cells of an invasive bladder tumor.
Vimentin expression. Novel expression of vimentin in bladder carcinoma cells in vivo was recorded in 143 of 795 (18%) tumors, including 29 superficial and 97 invasive lesions with 17 tumors that remained unstaged. Expression of vimentin (Fig. 1C and D) was significantly associated with tumor grade and stage (<0.001) but not with nodal involvement or vascular invasion. Figure 2 shows immunostaining for all markers in the same tissue core, demonstrating coexpression of the epithelial and mesenchymal intermediate filament proteins keratin (Fig. 2A) and vimentin (Fig. 2E), respectively.
Immunohistochemical staining of different sections from a single tissue microarray showing the same tumor core incubated with antibodies to keratin (A), E-cadherin (B), β-catenin (C), plakoglobin (D), vimentin (E), and N-cadherin (F). Note coexpression of intermediate filaments keratin and vimentin (compare A and E) in the absence of N-cadherin expression (F).
Associations among EMT markers. Loss or reduced expression of E-cadherin was significantly associated with low β-catenin (P < 0.01; Spearman correlation coefficient, 0.446), low plakoglobin (P < 0.01; Spearman correlation coefficient, 0.370), and novel expression of vimentin (P < 0.01; Spearman correlation coefficient, −0.109). Low expression of β-catenin was independently associated with low plakoglobin expression (P < 0.01; Spearman correlation coefficient, 0.441) and vimentin expression (P < 0.01; Spearman correlation coefficient, −0.181). In turn, low plakoglobin was found significantly associated with vimentin expression (P < 0.01; Spearman correlation coefficient, −0.231). No significant associations were found with any of the aforementioned EMT markers and N-cadherin.
Survival analysis. Because there was no complete follow-up data for the superficial group, survival analysis was confined to the cystectomy patient group. Univariate analysis involving each of the proposed EMT markers in overall survival showed that the reduction or loss of plakoglobin expression (P = 0.02) was significantly associated with patient survival with a trend for association with β-catenin (P = 0.11; Fig. 3A and C ). In contrast, no significant difference was recorded for E-cadherin or N-cadherin levels of expression (Fig. 3E and G). Within this patient group, the clinical variables of T-stage, lymph node status, and vascular invasion showed a significant correlation with survival, as previously established. The 5-year survival rate for patients whose tumors displayed a loss or reduction in expression of plakoglobin was 29.4% compared with 41.5% when plakoglobin displayed normal levels of expression. Ten-year survival rates were 22.9% and 29.2%, respectively, when the same plakoglobin variables were analyzed. The median time to death for patients with loss or reduced plakoglobin expression was 2 years compared with 4 years when a normal plakoglobin level was recorded. The 5-year survival rate for patients whose tumors displayed a loss or reduction in expression of β-catenin was 29.1% compared with 40.8% when β-catenin displayed normal levels of membrane staining. Ten-year survival rates were 24.2% and 23.5%, respectively, when the same β-catenin variables were analyzed. The median time to death for loss or reduced β-catenin was 2.4 years compared with 3.8 years when β-catenin displayed high expression.
Kaplan-Meier curves among patients with bladder cancer after radical cystectomy. Plakoglobin expression—overall (A) and cancer-specific (B) survival. β-Catenin expression—overall (C) and cancer-specific (D) survival. E-cadherin expression—overall (E) and cancer-specific (F) survival. N-cadherin expression—overall (G) and cancer-specific (H) survival, respectively.
When the outcome was limited to cancer-specific survival, the loss or reduction in plakoglobin expression (P = 0.02) and the reduction or loss of expression of β-catenin (P = 0.02) were significant for survival (Fig. 3B and D). As expected, T-stage, lymph node status, and vascular invasion correlated with cancer-specific survival. The 5- and 10-year survival rates were 43.7% and 41%, respectively, for loss or reduction of plakoglobin compared with 55% and 52.7%, respectively, in the group with normal levels of expression. With β-catenin, 5- and 10-year survival rates were 41.9% and 39.4%, respectively, when a loss or reduction of expression was recorded compared with 56.3% and 53.7%, respectively, for normal levels of expression. E-cadherin and N-cadherin did not show a significant difference for cancer-specific survival (Fig. 3F and H).
Multivariate analysis was done using plakoglobin, β-catenin, tumor stage, age, and vascular invasion because all showed significance in univariate analysis as recorded in Tables 4 and 5 . For overall survival, vascular invasion and age of diagnosis were strong independent prognostic predictors (hazard ratio; 1.83; P < 0.001 and hazard ratio; 1.03 per year increment, P = 0.006, respectively). Loss of plakoglobin had a hazard ratio of 1.31 (P = 0.11), and β-catenin was no longer associated with survival (P = 0.82).
Univariate Cox regression analysis
Cox proportional-hazards regression model—multivariate analysis
For cancer-specific survival, plakoglobin and β-catenin sites were included in the model with the aforementioned variables. Vascular invasion was the only significant independent predictor in the model (hazard ratio, 2.57; P < 0.001).
In this study, we have assessed the expression and prognostic value of a panel of five markers associated with the EMT phenotype. Loss of E-cadherin and novel expression of N-cadherin represent defining features of EMT, both of which have been reported in bladder tumorigenesis (22, 23). Linked to this transition has been the expression of the mesenchymal intermediate filament protein vimentin, often accompanied by altered cytokeratin expression (19). In addition, delocalization of β-catenin has been implicated in the EMT process (9). Numerous studies have shown loss or reduced expression of plakoglobin in epithelial cells in the absence of E-cadherin expression and more recently this phenotypic change has been implicated as part of the EMT phenotype (24). Changes in the aforementioned proteins represent EMT end points in which the expression of each may be modulated by a growing number of molecules involved in transcriptional regulation, cell signaling, and modulation of the tumor microenvironment (for review, see refs. 7, 14, 19, 24). However, there remains some controversy over the occurrence of such events in vivo originating from the apparent rarity of the EMT-like morphologic changes observed by pathologists in primary tumor sections and secondary lesions (8).
The counter argument to this necessarily requires recording EMT-related events in vivo during tumor progression (25, 26). Evidence for such events is beginning to emerge and presently include reports of Snail 1 protein, αvβ6 integrin, and nuclear β-catenin visualized at the invading edge of colon cancer tumors (14, 19). A search for the expression of such EMT-related proteins may be best served from the analysis of total tumor sections because the reported limited localization of proteins to the invasive front of the tumor may be missed in alternative tissue sampling formats.
A reduction or loss in expression of E-cadherin has long been recognized as an important primary event in bladder tumorigenesis often linked to poor prognosis (23, 27–29). In this study, altered E-cadherin expression was significantly associated with low β-catenin and plakoglobin expression along with novel expression of vimentin. Consideration of changes associated with the cadherin complex proteins revealed a significant difference between the frequency of such events in the superficial and invasive tumor groups, linking such changes to tumor progression. Although altered expression or localization of E-cadherin were not found associated with lymph node involvement or vascular invasion, the cytoplasmic localization of plakoglobin was a significant indicator of lymph node involvement.
Nuclear localization of β-catenin and plakoglobin was recorded in 3.8% and 1.8% of tumor samples, respectively, and was always observed in the presence of cytoplasmic localization of the catenin member. The reported frequency of nuclear localization of β-catenin in bladder tumors ranges between 0% and 22% identified in study groups involving small patient numbers (30–33). Mutations associated with β-catenin have been shown to result in the nuclear localization of this catenin family member in bladder tumors (30). Nuclear localization of plakoglobin seems to be a less frequent finding and although nuclear plakoglobin has been reported in other cancers (34, 35), we are not aware of any reports of this finding in bladder tumors. The nuclear localization of either β-catenin or plakoglobin was observed throughout the tumor section in a majority of samples whereby isolated islands of cells representing <5% of tumor cells with apparent nuclear staining were not scored as positive in this study. Given the potential putative transient nature of EMT and the limited, possibly temporary, nuclear relocalization of catenins associated with the invasive edge of the tumor, we may have overlooked more minor representation of EMT events.
Novel expression of N-cadherin has also been reported in human bladder tumors (22) and was previously recorded in 39% (20 of 51) of tumors. In this more extensive study, we have recorded N-cadherin expression in only 8.2% (61 of 746) of bladder tumors using the same antibody and antigen retrieval technique. Consistent with our previous report, novel expression of N-cadherin was observed to be focal in nature throughout the tissue microarray, and although the presence of four cores of each tissue is calculated to be representative of the tumor phenotype we believe that the N-cadherin expression recorded in this study is an underrepresentation of the frequency of expression in bladder tumors. Support for this interpretation is found in the scores for each of the four cores from a single tumor. A review of the results revealed detection of N-cadherin expression in all of the cores from a single tumor in approximately one third of the cases (21 of 61). It is also of interest that N-cadherin expression was recorded approximately equally in the superficial and invasive tumor groups. Moreover, in one recent publication, N-cadherin was identified as a prognostic marker of progression in superficial urothelial tumors (36). Although we have shown that novel expression of N-cadherin in bladder carcinoma cells promotes invasion (37), the expression of N-cadherin throughout different tumor grades and stage makes identification of additional functions a necessary consideration.
Novel vimentin expression, the second mesenchymal marker linked to EMT, was frequently recorded in the bladder carcinoma cell lines that displayed the more stellate, fibroblast morphology. However, past experience has revealed that normal urothelium can express vimentin after an extended period of in vitro culture (38). In this study, we have shown novel expression of vimentin in bladder carcinoma cells in vivo mainly associated with invasive bladder lesions. Confirmation of the epithelial origin of vimentin-positive cells was established in neighboring sections stained for total keratin demonstrating coexpression of these two intermediate filament types. No association was found between expression of the two mesenchymal markers, N-cadherin and vimentin. Whereas N-cadherin was found not to be associated with any of the clinical variables studied, vimentin expression was significantly associated with tumor grade and stage. In addition, novel vimentin expression showed significant association with reduced expression of both β-catenin and plakoglobin. Expression of vimentin was recorded in 18% (143 of 795) of bladder tumors, establishing novel expression of this intermediate filament in vivo. It is interesting that a subgroup of superficial tumors (6.8%) was identified that displayed novel expression of vimentin, where, given the association found with tumor stage and vimentin in invasive tumors, it is desirable to establish whether this marker identifies superficial tumors at heightened risk to progress as has been reported for N-cadherin (36).
In this study, survival data involved only patients that underwent cystectomy. For overall survival, the reduction or loss of plakoglobin was the one putative EMT variable that showed significant association with patient survival. As expected within this group of patients, the clinical variables of tumor stage, lymph node status, and vascular invasion also showed a correlation with survival. The median time to death for patients with loss or reduced plakoglobin expression was half that (2 years) recorded for patients with high expression of plakoglobin (4 years). Indeed, a previous study within a small group of bladder cancer patients (39) also reported that altered plakoglobin expression correlated with poor survival. In addition, alterations in plakoglobin expression have recently been identified as a marker of progression in T1 bladder tumors (40). A similar pattern of loss or reduced expression was recorded for β-catenin, although such differences did not reach statistical significance. When the outcome was limited to cancer-specific survival, both the expression level and delocalization of plakoglobin and β-catenin were found to have prognostic significance for survival.
In this study, we have identified changes linked to EMT in different models as events that occur in bladder tumors in vivo. Loss of E-cadherin in bladder tumorigenesis is well established and, more recently, we have shown novel expression of N-cadherin in human bladder tumors (22). However, reduced expression of plakoglobin associated with these cadherin events has only recently been linked to EMT (24). This combination of phenotypic changes has been observed in all bladder carcinoma cell lines that display the morphologic EMT phenotype accompanied by the enhancement of migration and invasion. In this study, we have shown that loss of E-cadherin expression is significantly associated with low plakoglobin expression even in the presence of N-cadherin expression. In addition, novel expression of vimentin was also found significantly associated with alterations of E-cadherin, β-catenin, and plakoglobin expression. The lack of association of N-cadherin with variables assessed as part of the EMT phenotype is surprising, although, as previously stated, we believe there is an underestimation of the frequency of novel expression of N-cadherin. However, despite this caveat, we have established the in vivo expression of putative EMT alterations in bladder cancer in which loss or reduced plakoglobin expression presented as a significant prognostic feature in these tumors.
We thank Barbara Rothman for the technical help in immunohistochemical staining of tissue microarrays, and Robert Kelley and Matthew Wszolek for help in collecting materials and data.
Grant support: R01-DK59400 (I.C. Summerhayes). B. Silva Neto and A. Biolo were sponsored by Coordenacao de Aperfeicoamento de Pessoal de Ensino Superior.
Note: E. Baumgart, M.S. Cohen, and B. Silva Neto contributed equally to this work and are listed in alphabetical order.
Received September 20, 2006.
Revision received December 18, 2006.
Accepted January 10, 2007.
Savanger P, Boyer B, Valles AM, Jouanneau J, Thiery JP. Modulations of the epithelial phenotype during embryogenesis and cancer progression. Cancer Treat Res 1994;71:229–49.
Hay ED. An overview of epithelio-mesenchymal transformation. Acta Anat 1995;154:8–20.
Shook D, Keller R. Mechanisms, mechanics and function of the epithelial-mesenchymal transition in early development. Mech Dev 2003;120:1351–83.
Thiery JP. Epithelial-mesenchymal transitions in tumor progression. Nat Rev Cancer 2002;2:442–54.
Thiery JP. Epithelial-mesenchymal transitions in development and pathologies. Curr Opin Cell Biol 2003;15:40–6.
Thompson EW, Newgreen DF. Carcinoma invasion and metastasis: a role for epithelial-mesenchymal transition? Cancer Res 2005;65:5991–5.
Huber M, Kraut N, Beug H. Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol 2005;17:548–58.
Tarin D. The fallacy of epithelial mesenchymal transition in neoplasia. Cancer Res 2005;65:5996–6001.
Brabletz TA, Jung S, Reu M, et al. Variable β-catenin expression in colorectal cancers indicates tumor progression driven by the tumor microenvironment. Proc Natl Acad Sci U S A 2001;98:10356–61.
Nakajima S, Doi R, Toyoda E, et al. N-cadherin expression and epithelial-mesenchymal transition in pancreatic carcinoma. Clin Cancer Res 2004;10:4125–33.
Bates RC. Colorectal cancer progression: integrin αvβ6 and the epithelial-mesenchymal transition (EMT). Cell Cycle 2005;4:1350–2.
Giroldi LA, Bringuier P-B, Shimazui T, Jansen K, Schalken JA. Changes in cadherin-catenin complexes in the progression of human bladder carcinoma. Int J Cancer 1999;82:70–6.
Miettinen PJ, Ebner R, Lopez AR, Derynck R. TGF-β induced transdifferentiation of mammary epithelial cells to mesenchymal cells: involvement of type I receptors. J Cell Biol 1994;127:2021–36.
Savagner P. Leaving the neighborhood: molecular mechanisms involved during epithelial-mesenchymal transition. Bioessays 2001;23:912–23.
Nawshad A, Lagamba D, Polad A, Hay ED. Transforming growth factor-β signaling during epithelial-mesenchymal transformation: implications for embryogenesis and tumor metastasis. Cells Tissues Organs 2005;179:11–23.
Bellovin DI, Bates RC, Muzikansky A, Rimm DL, Mercurio AM. Altered localization of p120 during epithelial to mesenchymal transition of colon carcinoma is prognostic for aggressive disease. Cancer Res 2005;65:10938–45.
Maeda M, Johnson KR, Wheelock MJ. Cadherin switching: essential for behavioral but not morphological changes during an epithelium-to-mesenchyme transition. J Cell Sci 2005;118:873–87.
Yang AD, Camp ER, Fan F, et al. Vascular endothelial growth factor receptor-1 activation mediates epithelial to mesenchymal transition in human pancreatic carcinoma cells. Cancer Res 2006;66:46–51.
Lee J, Dedhar S, Kalluri R, Thompson E. The epithelial-mesenchymal transition: new insights in signaling, development, and disease. J Cell Biol 2006;172:973–81.
Rieger-Christ KM, Ng L, Hanley RS, et al. Restoration of plakoglobin expression in bladder carcinoma cell lines suppresses cell migration and tumorigenic potential. Br J Cancer 2005;92:2153–9.
Gee J, Greathead P, Little AF, Libertino JA, Summerhayes IC. Aberrant cadherin expression in bladder carcinoma cells. Mol Urol 1998;2:73–81.
Rieger-Christ KM, Cain J, Braasch J, et al. Expression of classic cadherins type I in urothelial neoplastic progression. Hum Pathol 2001;32:18–23.
Shimazui T, Schalken JA, Giroldi LA, et al. Prognostic value of cadherin-associated molecules (α, β and γ-catenins and p120cas) in bladder tumors. Cancer Res 1996;56:4145–58.
Yang J, Mami SA, Donaher JL, et al. Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 2004;17:927–39.
Xue C, Plieth D, Venkov C, Xu C, Neilson EG. The gatekeeper effect of epithelial-mesenchymal transition regulates the frequency of breast cancer metastasis. Cancer Res 2003;63:3386–94.
Willipinski-Stapelfeldt B, Riethdorf S, Assmann V, et al. Changes in cytoskeletal protein composition indicative of an epithelial-mesenchymal transition in human micrometastatic and primary breast carcinoma cells. Clin Cancer Res 2005;11:8006–14.
Arance A, Fabra A, Germá JR. Prognostic value of the expression of E-cadherin and β-catenin in bladder cancer. Eur J Cancer 2000;36:357–62.
Miahle A, Louis J, Montlevier S, et al. Expression of E-cadherin and α-, β- and γ-catenin in human bladder carcinomas: are they good prognostic factors? Invasion Metastasis 1997;17:124–37.
Nakopoulou L, Zervas A, Gakiopoulou-Givalou H, et al. Prognostic value of the expression of E-cadherin and β-catenin in bladder cancer. Eur J Cancer 2000;36:357–62.
Shiina H, Igawa M, Shigeno K, et al. β-Catenin mutations correlate with over expression of c-myc and cyclin D1 genes in bladder cancer. J Urol 2002;168:2220–6.
Zhu X, Kanai Y, Saito A, Kondo Y, Hirohashi S. Aberrant expression of β-catenin and mutation of exon 3 of the β-catenin gene in renal and urothelial carcinomas. Pathol Int 2000;50:945–52.
Stoehr R, Kreig RC, Knuechel R, et al. No evidence for involvement of β-catenin and APC in urothelial carcinomas. Int J Oncol 2002;20:905–11.
Urakami S, Shiina H, Enokida H, et al. Epigenetic inactivation of Wnt inhibitory factor-1 plays an important role in bladder cancer through aberrant canonical Wnt/β-catenin signaling pathway. Clin Cancer Res 2006;12:383–91.
Aaltomaa S, Lipponen P, Karja V, Lundstedt S, Lappi J, Kosma VM. The expression and prognostic value of α-, β-, and γ-catenins in renal cell carcinoma. Anticancer Res 2004;24:2407–13.
Voutilainen KA, Anttila MA, Sillanpaa SM, et al. Prognostic significance of E-cadherin-catenin complex in epithelial ovarian cancer. J Clin Pathol 2006;59:460–7.
Lascombe I, Clairotte A, Fauconnet S, et al. N-cadherin as a novel prognostic marker of progression in superficial urothelial tumors. Clin Cancer Res 2006;12:2780–7.
Rieger-Christ KM, Lee P, Zagha R, et al. Novel expression of N-cadherin elicits in vitro bladder cell invasion via the Akt signaling pathway. Oncogene 2004;23:4745–53.
Summerhayes IC, Cheng YS, Sun TT, Chen LB. Expression of keratin and vimentin intermediate filaments in rabbit bladder epithelial cells at different stages of benzo[a]pyrene-induced neoplastic progression. J Cell Biol 1981;90:63–9.
Syrigos KN, Harrington K, Waxman J, Krausz T, Pignatelli M. Altered γ-Catenin expression correlates with poor survival in patients with bladder cancer. J Urol 1998;160:1889–93.
Clairotte A, Lascombe I, Fauconnet S, et al. Expression of E-cadherin and α-, β-, γ-catenins in patients with bladder cancer: identification of γ-catenin as a new prognostic marker of neoplastic progresion in T1 superficial urothelial tumors. Am J Clin Pathol 2006;125:119–26.
You are going to email the following Identification and Prognostic Significance of an Epithelial-Mesenchymal Transition Expression Profile in Human Bladder Tumors
Clin Cancer Res March 15 2007 (13) (6) 1685-1694; DOI: 10.1158/1078-0432.CCR-06-2330
Contact Guidance Controls T-cell Migration in PDAC
MET in Papillary RCC
Identification of T-cell Antigens by Minigene Screening
Show more Human Cancer Biology


text:
IT Conversations News: December 18, 2005
December 18, 2005 ~ Doug
Richard Monson-Haefel – The Rebel Platforms (rated 2.3 by our listeners) For enterprise application development, the high-end "superplatforms" like J2EE and .NET aren't the only choice. Developers can choose from the "rebel platforms," open-source platforms that don't adhere to industry standards like J2EE or .NET. Burton Group analyst Richard Monson-Haefel describes the rebel platforms, compares then with superplatforms from Microsoft, SAP, IBM, Oracle, and BEA on criteria including flexibility, risk, lock-in, development complexity, and cost.
Heather Gold (3.1) Moria Gunn interviews Heather Gold, high-tech humorist and creator of "I look like an egg, but I identify as a cookie." They'll talk about high-tech humor — yes, there actually is such a thing.
Mitchell Baker – The Mozilla Foundation (3.1) With over 100 million downloads so far, Firefox is a huge success. Mitchell Baker and her team helped blaze the trail for commercial open source. She shares her insights on The Mozilla Foundation as an organization and the launch of Firefox. From the O'Reilly Media Open Source Convention (OSCON).
Mary Roach (3.3) Moira Gunn also speaks with Mary Roach. Following her earlier New York Times bestseller "Stiff — The Curious Lives of Human Cadavers," Mary takes on another unusual subject: How Science studies the Afterlife. Mary will tell us what happened when she ran into Alyson Dubois, the real-life clairvoyant whose life is the basis for the hit show, Medium.
Robert Beardsley (3.3) Then on BioTech Nation, Moira speaks with Dr. Robert Beardsley, president and CEO of Kereos. He tells us about their new approach to detecting cancer.
Reinventing Media – Supernova 2005 (3.3) As the Internet becomes a greater factor in our day-to-day lives, the role of all forms of media is rapidly changing. Dan Gillmor leads a diverse group of entrepreneurs to discuss the evolution of the Internet from a platform for text and pictures into a platform for every sort of media.
Josh Bancroft – Podcasting from Mobile Devices (3.5) To produce great podcasts you don't need a lot of complex equipment or technical expertise. In this session from the first in-person Podcast Academy classes, Josh Bancroft tells how to get good results with a cellphone, PDA or MP3 player.
Ray Kurzweil – When Humans Transcend Biology (4.1) Increasing technological advances are certainly making a global impact. However, one may very well ask how new technologies will change daily life. In this question and answer session, Dr. Moira Gunn of Tech Nation talks with Ray Kurzweil about the impact of technological change on large and small arenas of life. This is part 2, the Q&A session. It's the unedited version of an earlier Tech Nation program.
Todd Kuiken and Jesse Sullivan – Mind and Body (too late for reviews) As director at the Neural Engineering Center for Artificial Limbs at the Rehabilitation Institute of Chicago, Dr. Todd Kuiken has found both a partner and a patient in Jesse Sullivan–a double amputee who has become the world's first bionic man. This presentation at Pop!Tech shows Jesse as he is: a remarkable man, possessing the patience of Job and a remarkable spirit.
This week's IT Conversations/O'Reilly Pick of the Week is a great one from early last year:
Marc Smith – Catalyzing Collective Action on the Net (3.8) This talk demonstrates several technologies and concepts that show promise as ways to enhance online communities, making them easier to discover and making it easier to select high-quality content, evaluate that content, and motivate others to contribute significant value. In short, these tools may catalyze collective action by highlighting participants' histories and relationships.
< Previous Update: The Conversations Network
Next > Interviewed by Eric Schwartzman

redpajama_set_name: RedPajamaCommonCrawl


text:
De La Mix Tape: Remixes, Rarities and Classics is a 2004 De La Soul compilation album from Rhino Records. It is similar to the A Tribe Called Quest compilation album Hits, Rarities & Remixes.

Personnel

Karen Ahmed – Project Assistant
Skeff Anselm – Producer
Vanessa Atkins – Project Assistant
Badmarsh & Shri – Remixing
Barry Benson – Compilation Producer
Joe Buck – Cover Design
Emily Cagan – Project Assistant
Chairman Mao – Liner Notes
Reggie Collins – Annotation
Pic Conley – Flute
Chris Conway – Mixing
DaBeatminerz – Producer
De La Soul – Arranger, Producer, Remixing, Mixing
DJ Honda – Producer
DJ Joc Max – Remixing
DJ Spinna – Remixing
Sue Faber – Mixing
Cory Frye – Project Assistant
Leigh Hall – Project Assistant
Dan Hersch – Remastering
Troy Hightower – Mixing
Jay Dee – Producer, Remixing
Masaki Koike – Art Direction, Design
Tim Latham – Mixing
Suzanne Love – Project Assistant
April Milek – Project Assistant
Mos Def – Vocals
Randy Perry – Project Assistant
Steve Pokorny – Remastering
Bob Power – Mixing
Prince Paul – Arranger, Producer, Mixing
Rockwilder – Producer
Gladys Sanchez – Project Assistant
Tim Scanlin – Project Assistant
Glenn Schwartz – Project Assistant
Dorothy Stefanski – Editorial Supervision
Truth Enola – Vocals
Diana Washburn – Project Assistant
Mason Williams – Project Assistant
Yankee B. – Vocals

References

De La Soul albums
Hip hop compilation albums
Albums produced by J Dilla
Albums produced by Prince Paul (producer)
Albums produced by Rockwilder
Albums produced by Da Beatminerz
2004 compilation albums
Hip hop remix albums
2004 remix albums
Rhino Records compilation albums
Rhino Records remix albums

redpajama_set_name: RedPajamaWikipedia


text:
Buy £30
The Burgundians: A Vanished Empire by Bart Van Loo, translated by Nancy Forest-Flier
A masterful history of the great dynasty of the Netherlands' Middle Ages.
'A sumptuous feast of a book' The Times, Books of the Year
'Thrillingly colourful and entertaining' Sunday Times
'A thrilling narrative of the brutal dazzlingly rich wildly ambitious duchy' Simon Sebag Montefiore
5 stars! Daily Telegraph
'A masterpiece' De Morgen
'A history book that reads like a thriller' Le Soir
At the end of the fifteenth century, Burgundy was extinguished as an independent state. It had been a fabulously wealthy, turbulent region situated between France and Germany, with close links to the English kingdom. Torn apart by the dynastic struggles of early modern Europe, this extraordinary realm vanished from the map. But it became the cradle of what we now know as the Low Countries, modern Belgium and the Netherlands. This is the story of a thousand years, a compulsively readable narrative history of ambitious aristocrats, family dysfunction, treachery, savage battles, luxury and madness. It is about the decline of knightly ideals and the awakening of individualism and of cities, the struggle for dominance in the heart of northern Europe, bloody military campaigns and fatally bad marriages. It is also a remarkable cultural history, of great art and architecture and music emerging despite the violence and the chaos of the tension between rival dynasties.
Head of Zeus, an Apollo book * Medieval History
28 Oct 2021 * 624pp * £30 * 9781789543438
'Bart Van Loo does something extremely difficult; he brings to life an illusion of a state in an unfamiliar world. And he does this with such verve and energy that you very nearly believe it'
'A thrilling narrative of the brutal dazzlingly rich wildly ambitious duchy that was the most advanced and sophisticated economy and the most extravagant flashy court of its time. Filled with flamboyant murderous and debauched dukes, courtesans, courtiers and maniacs, it is a total pleasure to read'
Simon Sebag Montefiore, Aspects of History, Books of the Year
'In this perky popular history, a bestseller on mainland Europe, Bart Van Loo traces the steady rise and sudden end of the Burgundians, sprinkling his narrative with many entertaining asides'
Bart van Loo is in top form. The Burgundians reads like a train and hits you like a sledgehammer. A masterpiece... He does not try to be a wise man or a moralist. What he does make clear is that that the history of early Dutch unification is one that came about both through excessive bloodshed and praiseworthy magnanimity. Mission accomplished'
'History told – and well told, too – for those who value narrative at least as much as the finicky details of economics or treaty-making'
To narrate the legendary story of the dukes of Burgundy, you need a learned and visionary guide like Bart Van Loo... A masterful work'
'Suitably epic'
BBC History Magazine
A sparkling history of the origins of the Low Countries... Van Loo arouses interest in the past among thousands of readers, spectators and listeners in an inimitable way'
The Low Countries
'A pleasure to read from start to finish. How fortunate that Van Loo is not just a historian but also a writer. Truly spectacular!'
Neue Züricher Zeitung
The political and the personal, economics and culture, belief and violence, success and failure, major developments and spicy details – it's all there. The Burgundians expertly draws on the latest scientific insights, but is also told with lightness and elegance'
Frits van Oostrom
'The formidable saga of our Burgundian origins. Over 650 pages that read like a great political adventure novel, a Game of Thrones soap opera where everything is true'
La Libre Belgique
'Full of cliff hangers and moving passages. Irresistible'
Herman Pleij
'Bart van Loo is back and emerges once again as a true storyteller. Van Loo is the perfect guide through the past. It is as if we are there'
'A history book that reads like a thriller'
'Colourful and multidimensional: a Belgian master storyteller'
Kulturradio, SWR2
Narrative history of the highest level... The author conjures the tastes, smells, colours and feelings of the past'
Kulturradio, WDR 3
'Thrillingly colourful and entertaining'
'Most of us think of wine when we hear the word 'Burgundy', but Bart Van Loo uncovers a lost empire of mad dukes, strange delicacies and great wealth'
The Times, Books of the Year
'A worthwhile and satisfying read'
A sumptuous feast of a book... Van Loo recreates the world of Ghent and Bruges in loving detail, a bustling, blood-soaked landscape of quays, merchants and money changers'
The Times, '21 best history books of 2021'
'Lively, anecdotal unpicking of this fascinating but nebulous entity'
'Belgian historian Bart Van Loo tells its story in 'rollicking' style'
'Stuffed with elaborate feasts and bloody battles, Van Loo's thrillingly colourful and entertaining book has been an enormous success in his native Belgium and it's easy to see why'
Bart Van Loo
Bart Van Loo has developed a rare twin talent over the years. While drawing big crowds in the theatre, he is also the author of the universally praised France Trilogy and the bestseller Chanson: A sung history of France. The Burgundians is the first of his books to be translated into English.
Nancy Forest-Flier
Nancy Forest-Flier grew up in a Dutch-American milieu in the United States and studied English literature and creative writing at Hope College, Michigan before settling in the Netherlands in 1982. She works as a freelance English-language editor and Dutch-English translator.
OTHER BOOKS YOU MIGHT LIKE
The Lost Chapel of Westminster


Our aim is to get your skin looking the best it possibly can.
Our experienced skin care professionals will work out the best treatment regime for you for in salon care and take home care.
Like a berocca for your skin, this quick pick-me-up is for when time is precious.
Includes a deep cleanse, skin analysis, exfoliation and steam, mask and beautiful scalp massage.
Specially selected for your skin type-our skin care professional will tailor make a facial just for you using the Environ range.
The aim of the treatment it to help improve signs of ageing,scaring, pigmentation, dehydration or problem skin.
This facial includes a deep cleanse, skin analysis, exfoliation and steam, personalised active serum, decolletage and facial massage, rubberised mask and scalp massage.
You receive finishing products for your skin.


Kapa'a, meaning "the solid or closed", lies within the ancient 'ili land division of Ka 'Ōhi'a, which translates to "the 'ōhi'a lehua trees are tabooed." Ka 'ōhi'a falls within the larger 'ahupua'a land division of Kailua. To the north [of Kapa'a] is Kawai Nui. The vast expanse of Kawai Nui was once rich with the delectably fat "mudfish": 'o'opu ku'ia. The 'o'opu was lured by the famous mythological Mākālei tree, the supernatural fish-attracting tree transplanted by the ancients from Pali'uli (Hilo). The broad silent waters of Kawai Nui were once guarded by the mo'o goddess, Hauwahine.
Standing mauka above Kawai Nui in the misty distance is pleasant Kapa'a, revered land of legendary ruler, Olopana.1 The northwestern boundary is a ridgeline known as MahinuiA, separating it from the Kawa Stream watershed of Kāne'ohe ahupua'a. The southern flank of Kapa'a Valley is the hilly place known as UlumawaoA.
The original "modern" Kapa'a Quarry (begun in late 19494) occupies the former site of the ancient stone adze quarry of the same name. The Hawaiian civilization depended upon the stone adze as a construction tool to produce religious images, build homes and houses of worship, produce agricultural tools, manufacture weapons of war, and carve and shape the all-important fishing canoes and the impressively large double-hulled voyaging canoes which attained lengths of 120 feet and more.1 The first modern quarry in Kapa'a Valley was filled by the City & County municipal waste landfill known as Kapa'a Landfill, operated from 1970 to May 6, 19972. Stone pieces—adze blanks—were transported to villages around Kawai Nui marsh for shaping. Numerous grind stones can be observed near the marsh shore today, where chips and blanks are regularly foundA.
Looking into Kapa'a Valley from the vicinity of the Mokapu Saddle Road. A small portion of Kawai Nui Marsh and the Kapaa Quarry Road are evident beyond H-3 on the left. The now-closed City & County Kapa' Landfill occupies the center of the photograph.
Because of the H-3 Interstate Highway, Kapa'a is familiar to most windward residents. This highway passes from Kāne'ohe to Mōkapu through Kapa'a Valley, at the upper end through the saddle between Ulumawao and MahinuiB near Hawaii Pacific University. This four-lane freeway now dominates the upper part of Kapa'a Valley, comprising some 13 acres (5.3 ha) or 2% of the watershed. The highway divides much of the area between the two principal land uses: the Ameron rock quarry operation on one side (Photographs 2, 3, & 4) and the City & County solid waste facilities on the other (Photograph 1). But the division is not perfect. In the upper valley, Ameron has developed its Phase II quarry operation on the slopes of Ulumawao across the freeway from now abandoned Phase I.
Looking towards the head end of Kapa'a Valley, with one of the peaks of Ulumawao on the left, the H-3 where it enters the upper end of Kapa'a Valley, and part of the Ameron quarry operation on the right.
1972. The western part, in the valley, was the Kalaheo Landfill from 1986 to 1990; then became a commercial green-waste facility. A model airplane park occupies a portion of the former landfill placed in Kawai Nui2,3.
Vertical view looking down into the Ameron quarry (Phase I pit) in Kapa'a Valley. The forested slope below the ridgeline at the bottom of the photograph is part of the watershed of Kawa Stream.
Bottom of the Ameron quarry (Phase I) operation in Kapa'a. This quarry pit was the source of most of the construction aggregate used on O'ahu. Abandoned as a source of construction material, it is now a lake.
1 Modified from "Cultural History of Kapa`a Quarry" written by Palani Vaughan for Ameron HC&D, May 18, 1993 (xerographic).
2 State of Hawaii Department of health, Solid Waste Section, Landfill Database - Oahu at CLICK HERE.
3 Young, R. 2005. Garbage in Paradise: A History of Honolulu's Refuse Division . Division of Refuse Collection, Dept. of Environmental Services.
4 HDOH. 2007. Total Maximum Daily Loads (TMDLs) for Total Suspended Solids, Nitrogen and Phosphorus in Kapa'a Stream, Kailua, Hawaii .
A It is possible to observe evidence of this commerece in tool-making at an ancient village site ("Ka Lā" ?) at Nā Pōhaku State Park Reserve off Kapa'a Quarry Road.
used in earlier USGS maps for just Ulumawao.


text:
Business, Arts & Culture
Dreamcatcher business catching on, expands to SK
Jeanelle Mandes | January 06, 2015
Dennis Cameron and his wife Michelle recently celebrated the grand opening of their Regina office for their business Dreamcatcher Promotions.
Since forming their business in 2012, they have created a head office in Tootinaowaziibeeng Treaty Reserve and also have a showroom in Winnipeg. They decided to branch out into Saskatchewan to expand their opportunity. "We want to show people that it can be done, you can succeed and own your own business," said Cameron, a former Chief.
Dreamcatcher Promotions does customized embroidery, screen printing, and full printing services for calendars. "We pretty much do the decorating of apparel such as jackets, t-shirts, hoodies, tracksuits, hats, scarves and a wide variety of clothes, we can customize it," says Cameron. "We also do promotional products such as pens, cups, lanyards, and frisbees and pretty much anything that corporate industries want to use to promote itself."
The couple both had the dream of owning their own business and helping the youth. Cameron says they also customize athletic uniforms of all types from hockey to baseball including uniforms in the North American Indigenous Games (NAIG) held this past summer in Regina.
"I'm proud to say we were one of the corporate sponsors... we donated 5000 of the athletes bags. We did a variety of the merchandise apparel they were selling during the games," says Cameron.
He says one of his business philosophies is to give back to the communities because that is how he was raised and to respect tradition.
"Some of my competitors think I'm crazy for doing that but it's just who I am. I know some of the obstacles that the communities have to deal with," he adds.
Cameron says what sets them apart from other companies is their pricing. He says they are more considerate and aware of the situations on First Nation communities with budget restraints.
"I'm in a position to help accommodate and ensure that our young kids are dressed nice and have that full apparel," he says. "I remember growing up myself; I looked like a Christmas tree. I had different coloured socks and we had the old jerseys. Everybody else came out looking all nice and we're awe wish we could've be like that," he says laughing.
This past Christmas, they offered a winter special and put together a winter package at an affordable price. The couple hopes to expand their business 'Dreamcatcher Promotions' to Saskatoon and Prince Albert to create employment for others.
Click here for more Arts & Culture stories and here for more Business stories.
< Back to EFN Business


I was first introduced to Aesop by my sister. I am known for my tendency to get extremely dry skin and I am also known for not doing anything about it. So, one Christmas, she gifted me the Aesop hand balm. And a good present it was. This hand balm moisturises so well and for so long. That hand balm also made me curious to try some of Aesop's other products.
I used to have a theory that Asian facial moisturisers were the best for my skin. This was because I had already spent hundreds of dollars on expensive designer creams and it didn't seem like they ever did anything for me. When I visited Hong Kong and discovered places like Watson's and Sasa, I went crazy! Skincare was so cheap and there was such a variety. I bought a cream that cost me about AUD$10 and it worked wonders. I can't tell you want brand it was or what it's purpose was, because, to be quite honest, I didn't understand a word that was written on the packaging. And that language barrier made it really difficult to get more of that same product when I returned back to Sydney. I spent years trialing all different sorts of moisturisers until I found Aesop. For my face, I use their Parsley Seed Anti-Oxidant Hydrator, which, I am told, is great for Australia's climate. It is really light and makes me feel fresh - perfect for my skin-type (normal to oily).
After two wins by Aesop, I was keen to try another product. I noticed my hair salon's bathroom always had an Aesop hand wash that contained exfoliating beads. Now, some of you might be thinking, "Exfoliating hand wash? What's new?". Well, for me, I had never seen or used one before and it amazed me. Therefore, my next Aesop investment was their Reverence Aromatique Hand Wash.
So, that's three wins by Aesop! The prices are a little on the higher side, but unlike some of those expensive designer skincare products, I find that I'm actually getting what I paid for.
Shop Navy Blue Blouses Tops, Black Heels, Black Chokers, Long Dresses and more. Get this widget.


text:
⇑ back to "Church & Ministry"
The Reconnection Mandate Directive No. 3: Pursue True Repentance
4:00PM EDT 10/12/2020	 Grant Berry and Leslie Crincoli
Note: This is the third in a series of five articles. Click here for directives No. 1 and No. 2. This document that was given from the Lord to Grant Berry to help the body of Messiah/Christ move into restoration in the family of God.
Repentance Defined
For restoration to be realized in "the One New Man" (TONM) between believing Jews and Gentiles, true repentance is necessary. First, let's define repentance, and second, understand its application in the context of the reconnection message.
Merriam-Webster's Dictionary defines repentance as "the action or process of repenting, especially for misdeeds or moral shortcomings." Repenting means "to turn from sin and dedicate oneself to the amendment of one's life." I sometimes think the modern church has lost the true meaning of this foundational tenet of the faith. It is not just about confession; confession is only the beginning of the process of repentance. If we have any doubts about the application of repentance, the apostle Paul brings a very clear explanation in Acts when speaking with King Agrippa: "I preached that they should repent and turn to God and demonstrate their repentance by their deeds" (Acts 26:20b, NIV). Repentance requires action to correct what is wrong.
Get Spirit-filled content delivered right to your inbox! Click here to subscribe to our newsletter.
Our understanding of the complete meaning of repentance is highly significant when it comes to the reconnection and moving into restoration in TONM. We need to grasp the full measure of our Father's directives and grant Him unlimited access to our hearts through which love and unity can thrive. We are at the time in history where Israel is awakening to her calling and destiny, and Jewish people are finding their identity in Yeshua/Jesus.
And so, God's children from the nations need to be fully reconciled with Israel's remnant (Jewish believers in Yeshua), and Israel's remnant needs to be fully reconciled with God's children from the nations. As we have already discussed in directives 1 and 2, this is an essential part of God's end-time plans to restore love and unity to the balance in His family. Full repentance here means that the Gentile branches wholly embrace the Jewish branches of the olive tree and the covenants and election of the people of Israel along with their own. One New Man does not mean one less Jew! Jewish identity needs to be restored among Jewish believers in the church/ekklesia, who must dwell in love unity with their non-Jewish family.
This Will Require a Shift
The reconnection will require a significant shift for all family members, especially the church, which has been separated from Israel, and its Jewish roots and heritage for over 1700 years. In this light, it is critical for the body of Messiah/Christ to recognize and address the personal issues that are keeping us separated, particularly the spiritual, emotional and theological barriers that have blocked harmony and unity between Messianic Jews and believing Gentiles. Parts of the church have acknowledged some issues, but not all of them, and the unity between Jewish and Gentile believers remains mostly fragmented and is still much misunderstood.
In Romans 12, Paul speaks of the heart transformation that revolutionizes relationships and challenges believers to live as one family. The essence of this transformation is based on repentance, mutual submission, humility and honesty on the parts of Jew and Gentile alike (see Chapters 2, 7 and 8 in Romans 9). Through the reconnection and alignment process, we understand that judgment begins in the house of God. In this context, judgment is not an adverse thing; rather, it is an act of mercy and healing from God.
The God of mercy wants to purify, heal and restore His family to one another (see Rom. 11:32), expose Satan's influences in these areas, and resolve these issues so that together as TONM, we gain authority to fulfill God's end-time plans on the earth. The body must come to understand the fullness of Satan's deceptions and be willing to uncover and eradicate them. We need to face them, or they will prevent us from embracing this reunion.
The Obstacles
We defined replacement theology in directive No. 2. Anti-Semitism is outward discrimination or acts of hostility towards and against Jewish people. Generational anti-Semitism is subtler; it is the influence of sin in our forefathers that travels from one generation to the next. Its effects are evident in much of the church, yet most are unaware of its influence. For example, a believer may know and believe there is still the promise for the Jewish people, yet their hearts can be stony and indifferent towards them.
This is because of generational anti-Semitism. The enemy has also stealthily woven the replacement mindset into the Gentile believing church to the degree that most are unaware of its influence. And it has spawned an identity in the church apart from Israel that is disconnected from its Jewish roots and heritage. The global church, planted by the Western church, has also been influenced in this way. So all of God's children from the nations need to examine themselves carefully when addressing the reconnection in TONM. The church/ekklesia must rid itself of all anti-Semitism, and the counterfeit belief of replacement theology in all its variations must be cut at the roots.
5 Directives for the Reconnection Mandate Between Christians and Messianic Jews, Part 2
5 Directives for the Reconnection Mandate Between Christians and Messianic Jews
The entire church must also be willing to confess and repent of any emotional issues such as jealousy or hatred, judgment or criticism that may be in our hearts and enter into a renewed love and support for our Jewish brethren. Likewise, Jewish believers need to address any heart issues that keep them apart and separate from loving their Gentile-believing family. These are the sibling obstacles that we need to ask God to remove from our hearts.
Most of these issues were seeded and fueled by the enemy as the God's children from the nations assumed control over the church (third and fourth centuries). As a result, these influences and flawed theology have traveled the generations to the modern day, mostly without repentance. We must point out that we are not personally responsible for the sins of our fathers and mothers.
However, erroneous doctrine or negative spiritual influences may still linger through the generational bloodline, affecting our minds and hearts, the way we think and act towards the Jewish people and Israel or even causing us to sin. Therefore, shouldn't we want to address all of these issues to get free of them?
Mending The Breach
Mending this breach between Jewish and Gentile believers helps pave the way to greater unity in the body of Messiah/Christ and restore the church to its former glory and move it into the "latter glory." A restored One New Man will empower the church to mend other divisions in the family of God (racial, theological and so forth), bringing it to wholeness and effectiveness. But first, we must come to this place of repentance.
To paraphrase Romans 12:2 in this light: Do not conform any longer to the old wineskin where Jew and Gentile were still separated in the One New Man, but be transformed into a new wineskin by the renewing of your mind. Then you will be able to test and approve what God's will is—His good, pleasing and perfect will.
Pursuing true repentance through the reconnection will prepare our hearts and minds to be renewed in the One New Man.
Please click here to watch the teaching and prayer over the 10 Days of Awe for Directive No. 3.
Author/speaker Grant Berry, founder of Reconnecting Ministries and Messiah's House Messianic Center, trumpets his new book, Romans 911—Time to Sound the Alarm! Grant is a Messianic believer in Yeshua, Jesus, and is promoting a new message into the body of Messiah, Christ, during these last days, known as "the reconnection." This is a new term used to describe the reuniting and full restoration of Jewish and Gentile believers into John 17 unity into the one new man now that Israel's awakening is upon us. This spiritual transaction has been unseen during most of the church age but is now most vital for the church to move into for God's end-time plans. It is a power key to the last great revival and final outpouring of the Holy Spirit on the earth. For more information and a special offer on Romans 911, please visit reconnectingministries.org.
Great Resources to help you excel in 2019! #1 John Eckhardt's "Prayers That..." 6-Book Bundle. Prayer helps you overcome anything life throws at you. Get a FREE Bonus with this bundle. #2 Learn to walk in the fullness of your purpose and destiny by living each day with Holy Spirit. Buy a set of Life in the Spirit, get a second set FREE.
Charisma Magazine
See an error in this article? Send us a correction
RELATED TOPICS: Grant Berry | Holy Spirit | Jesus | Jews | Reconnection | Repentance
Please follow these guidelines before commenting on our website:
Please be considerate and respectful of your fellow posters. If Jesus and your mother would not approve of your writing, you should revise your comment before submitting.
Use standard writing style and punctuation. Complete sentences and proper grammar will help others understand you.
Do not use profanity, obscenities, abusive language or otherwise objectionable content (as determined by Charisma Media moderators, in their sole discretion). Comments should not harass, abuse or threaten another's personal safety or property, make false statements, defame or impersonate someone else.
Do not question others' faith just because you disagree with them. It adds nothing to the discussion and only causes strife.
Do not question why someone comes to this website just because they disagree with you or the content of the article.
Do not post personal information including phone numbers, email or mailing addresses, or credit card information belonging to you or others.
Do not include copyrighted content or other intellectual property that you do not own or have the explicit rights or lisenses to distribute.
Do not post, directly or through links, viruses, misleading, malicious or off-topic content.
Do not use our comments section to advertise your website(s). This includes fequently linking to your site at the end your comments.
We reserve the right to deny commenting privileges of anyone for any period of time for any reason.
Please consider the following statements pertaining to comments posted by you and other visitors to our website:
Appearance of comments, advertisements or hyperlinks made by other commenters on this site do not necessarily indicate or constitute acceptance of or endorsement of the products, companies, corporations, ministries, organizations or agencies in whole or in part by staff members and/or the members of the editorial board of Charisma Media.
Comments are not pre-screened before they post. Charisma Media reserves the right to modify or remove any comment that does not comply with the above guidelines and to deny access of your Disqus account to make additional comments to the website without any notice. If you have been denied access to comment due to a violation of these terms please do not create multiple accounts in an attempt to circumvent the system. The correct course of action is to request a review of your account status by contacting webmaster@charismamedia.com.
Charisma Media is not responsible or liable in any way for comments posted by its users.
If you believe a comment is in violation of the above guidelines, U.S. law or International treaties, or is legitimate "trolling" please flag the post or contact webmaster@charismamedia.com. Include a link to the comment, along with a statement explaining what you believe the violation is and any evidence backing your claim. Charisma Media moderators will consider your request and decide on the most appropriate action.
View/Add Comments
Heaven is So Real


text:
Cybercrime | News | Privacy
Researchers go hunting for Netflix's Bandersnatch
Posted: March 22, 2019 by Christopher Boyd
New research from the Indian Institute of Technology Madras explains how popular Netflix interactive show Bandersnatch could fall victim to a side-channel attack and expose users' choices.
A new research paper from the Indian Institute of Technology Madras explains how popular Netflix interactive show Bandersnatch could fall victim to a side-channel attack.
In 2016, Netflix began adding TLS (Transport Layer Security) to their video content to ensure strangers couldn't eavesdrop on viewer habits. Essentially, now the videos on Netflix are hidden away behind HTTPS—encrypted and compressed.
Previously, Netflix had run into some optimisation issues when trialling the new security boost, but they got there in the end—which is great for subscribers. However, this new research illustrates that even with such measures in place, snoopers can still make accurate observations about their targets.
What is Bandersnatch?
Bandersnatch is a 2018 film on Netflix that is part of the science fiction series Black Mirror, an anthology about the ways technology can have unforeseen consequences. Bandersnatch gives viewers a choose-your-own-adventure-style experience, allowing for various options to perform task X or Y. Not all of them are important, but you'll never quite be sure what will steer you to one of 10 endings.
Charlie Brooker, the brains behind Bandersnatch and Black Mirror, was entirely aware of the new, incredibly popular wave of full motion video (FMV) games on platforms such as Steam [1], [2], [3]. Familiarity with Scott Adams text adventures and the choose your own adventure books of the '70s and '80s would also be a given.
No surprise, then, that Bandersnatch—essentially an interactive FMV game as a movie—became a smash hit. Also notable, continuing the video game link: It was built using Twine, a common method for piecing together interactive fiction in gaming circles.
Researchers figured out a way to determine which options were selected in any given play-through across multiple network environments. Browsers, networks, operating systems, connection type, and more were changed for 100 people during testing.
Bandersnatch offers two choices at multiple places throughout the story. There's a 10-second window to make that choice. If nothing is selected, it defaults to one of the options and continues on.
Under the hood, Bandersnatch is divided into multiple pieces, like a flowchart. Larger, overarching slices of script go about their business, while within those slices are smaller fragments where storyline can potentially branch out.
This is where we take a quick commercial break and introduce ourselves to JSON.
Who is JSON?
He won't be joining us. However, JavaScript Object Notation will.
Put simply, JSON is an easily-readable method of sending data between servers and web applications. In fact, it more closely resembles a notepad file than a pile of obscure code.
In Bandersnatch, there are a set of answers considered to be the default flow of the story. That data is prefetched, allowing users who choose the default or do nothing to stream continuously.
When a viewer reaches the point in the story where they must make a choice, a JSON file is triggered from the browser to let the Netflix server know. Do nothing in the 10-second window? Under the hood, the prefetched data continues to stream, and viewers continue their journey with the default storyline.
If the viewer chooses the other, non-default option, however, then the prefetched data is abandoned and a second, different type of JSON file is sent out requesting the alternate story path.
What we have here is a tale of two JSONs.
Although the traffic between the Netflix browser and its servers is encrypted, researchers in this latest study were able to decipher which choices its participants made 96 percent of the time by determining the number and type of JSON files sent.
Should we be worried?
This may not be a particularly big problem for Netflix viewers, yet. However, if threat actors could intercept and follow user choices using a similar side channel, they could build reasonable behavioral profiles of their victims.
For instance, viewers of Bandersnatch are asked questions like "Frosties or sugar-puffs?", "Visit therapist or follow Colin?", and "Throw tea over computer or shout at dad?". The choices made could potentially reveal benign information, such as food and music preferences, or more sensitive intel, such as a penchant for violence or political leanings.
Just as we can't second guess everyone's threat model (even for Netflix viewers), we also shouldn't dismiss this. There are plenty of dangerous ways monitoring along these lines could be abused, whether the data is SSL or not. Additionally, this is something most of us going about our business probably haven't accounted for, much less know what to do about it.
What we do know is that it's important that content providers—such as gaming studios or streaming services—affected by this research account for it, and look at ways of obfuscating data still further.
Afterall, a world where your supposedly private choices are actually parseable feels very much like a Black Mirror episode waiting to happen.
Christopher Boyd
Lead Malware Intelligence Analyst
Former Director of Research at FaceTime Security Labs. He has a very particular set of skills. Skills that make him a nightmare for threats like you.


text:
Q: Reactjs - create factory, passing props into component Trying to achieve the following:
if(typeof window === 'undefined'){
    return React.renderToString(React.createFactory(AppComponent)());
}
React.render(<AppComponent component={component} />, document.body);

How exactly can I pass in the component prop to the component when it's being put through the createFactory method?
EDIT:
Got it working:
renderApp: function(component){
    if(typeof window === 'undefined'){
        var appComponentFactory = React.createFactory(AppComponent);
        return React.renderToString(appComponentFactory({
            component: component
        }));
    }
    React.render(<AppComponent component={component} />, document.body);
}

A: If you're already transpiling JSX for the React.render() call, you don't need to jump through hoops with factories...
renderApp: function(component){
    var app = <AppComponent component={component}/>
    if (typeof window === 'undefined') {
      return React.renderToString(app);
    }
    React.render(app, document.body);
}

...since it's just sugar for:
var app = React.createElement(AppComponent, {component: component})

This may not apply, but If your AppComponent isn't doing anything but rendering the component prop it's given, you could also just render the component directly:
renderApp: function(Component) {
    var app = <Component/>
    if (typeof window === 'undefined') {
      return React.renderToString(app);
    }
    React.render(app, document.body);
}

...showing the desugaring again:
var app = React.createElement(Component)

redpajama_set_name: RedPajamaStackExchange


text:
Q: Any Chess AI for Flash? I am looking for any Chess AI for flash CS5 ActionScript 3.

A: A quick Google search reveals much of what you're looking for, including good discussions on the basic algorithms involved. If all you're looking for is a solution you can plug into your Chess game, don't expect anything terribly robust, especially if you're looking to outsmart DeepFritz or Rybka.
Should you be interested in the process of programming a Chess AI, you might want to read through the Chess Programming Wiki.
(As Drknezz points out, http://www.gnu.org/software/chess/ is also an excellent point of research for anyone interested in chess engines.)

A: You can check out my open source chess game project at Github:https://github.com/yangboz/godpaper


text:
What next for the Knicks?
Kristaps Porzingis by Kieth Allison is licensed under CC by 2.0
The New York Knicks are a team that have improved this season. This time last year they finished the season with an abysmal record of 17-65, finishing 14th in the Eastern Conference, only ahead of Philadelphia.
This year, the record looks a little more respectable. With 32 wins and 48 losses with 2 games remaining, there has been improvement. However, the season is yet again ending on a disappointing note, because at one stage the Knicks had an even 22-22 record. Since then they have only managed 1o wins, and lost 26.
Head Coach Derek Fisher was fired in early February, and assistant coach Kurt Rambis took charge until the end of the season, and has done an equally as bad, if not worse job, than Fisher. This week there have been rumours emerging around the league that Knicks president Phil Jackson wants to leave his role and return to the Lakers.
With no draft picks this summer, no permanent head coach, and potentially losing the President of the association, things look like they are beginning to fall apart in New York. So where do they go from here?
Well the most important factor in an organisation is of course the players. One of the Knicks' few successes this season was drafting 7'3″ Latvian Kristaps Porzingis. He has proved to be a huge talent, and has a lot of time to continue to improve. They also traded Tim Hardaway Jr to the Atlanta Hawks for the 16th pick in the draft last season, where they drafted point guard Jerian Grant from Notre Dame.
Robin Lopez has proved to be the best free agency acquisition for New York, and Knicks fans will have been pleased to see the Center put up some big points this season, taking the pressure off of Carmelo Anthony. Derrick Williams has made the NBA top 10 plays more than his fair share of times this season with some huge dunks, providing energy and points production coming off the bench.
Jose Calderon has failed to impress as the starting point guard, and the Knicks fans have shown their desire to see him replaced moving into next season. Arron Afflalo has had positive moments at shooting guard, but rumors have spread of his unhappiness in New York, and the video of him refusing to high-five teammate Kyle O'Quinn went viral, which didn't help his cause. He was moved to the second unit in an attempt to bring some scoring firepower to the bench, but this move has provided unsuccessful. Sasha Vujacic has proved to be very inconsistent, and the rest of the bench have had their good and bad moments.
I highly expect second unit members such as Langston Galloway and Kevin Seraphin to be in a Knicks uniform again next season, as well as Derrick Williams. I am expecting some members of the bench to be offloaded however, and I'm sure there will be some changes to the roster over the summer, and will look different next season. Maybe not a complete roster makeover like the 2015 offseason, but change can be expected.
Of course there is Carmelo Anthony, who has led the Knicks in points (21.9), assists (4.2) and rebounds (7.8). Anthony has been the core this season to whom the rest of the team have relied on to score points efficiently.
In my eyes, 3 of next seasons starting 5 are already decided. I expect Carmelo Anthony, Kristapz Porzingis, and Robin Lopez to be starting at the beginning of next season. So that of course leaves the backcourt, where there has been a lack of points production, and in my eyes is where it has all gone wrong.
For the second half of this season, the Knicks have been poor. Knowing that the playoffs were out of reach, they should have given the young guys more opportunities to start games. On friday night, rookie Point Guard Jerian Grant started the game against Philadelphia and had a great game. He scored 16 points, and also had 6 assists and 5 rebounds, playing a big part in the 109-102 victory.
Last week he also started against Brooklyn, and had 14 points and 4 assists in the 105-91 victory. These performances have both resulted in victories for the Knicks, and although they are against weak teams, it still shows that with some more game time, Grant has the potential to be a very good point guard for the Knicks. In my opinion, Coach Rambis should have given up on Jose Calderon a long time ago, and given Grant more opportunities as the season has gone on.
So with 2 games left in the season, and no post-season basketball in New York for yet another year, the future looks uncertain. Names have floated around the league as potential Head Coaches of the Knicks next season, but there continues to be no outright favorite for the role. With no draft picks in this years highly anticipated NBA draft either, the Knicks may well have to rely on free agency in order to bring any joy to the fans in NYC.
Unfortunately, no big names have been linked with New York for this upcoming free agency. With the current uncertainty and no picks going forward to build a future of young talent, big names are turning their heads away from the Big Apple. Mike Conley is the only free agent who has been linked with a switch to the Knicks, however most arrows are pointing elsewhere for the superstars of the draft.
Not all hope is lost however for the Knicks. We saw last year that Phil Jackson isn't afraid to trade away players for draft picks. So perhaps he has a plan to offload a player or two in order to grab a top 20 pick in this years draft.
Porzingis was the first step to the rebuilding phase of this New York franchise, now they need to take another big step forward this summer. Which step they take however, remains very unclear.
NBA All-Star Game Starters Announced
Long Live King James (but Kyrie is the MVP)


text:
#ifdef CONFIG_NET
int toi_prepare_usm(void);
void toi_cleanup_usm(void);

int toi_activate_storage(int force);
int toi_deactivate_storage(int force);
extern int toi_usm_init(void);
extern void toi_usm_exit(void);
#else
static inline int toi_usm_init(void)
{
	return 0;
}

static inline void toi_usm_exit(void)
{
}

static inline int toi_activate_storage(int force)
{
	return 0;
}

static inline int toi_deactivate_storage(int force)
{
	return 0;
}

static inline int toi_prepare_usm(void)
{
	return 0;
}

static inline void toi_cleanup_usm(void)
{
}
#endif

enum {
	USM_MSG_BASE = 0x10,

	/* Kernel -> Userspace */
	USM_MSG_CONNECT = 0x30,
	USM_MSG_DISCONNECT = 0x31,
	USM_MSG_SUCCESS = 0x40,
	USM_MSG_FAILED = 0x41,

	USM_MSG_MAX,
};

redpajama_set_name: RedPajamaGithub


St. Ignatius believed that you can find evidence of God in all things. I believe it, too … and here's how I've been finding God lately.
and Minerva Lake in the Adirondacks, where Scott's cousin organized a family reunion.
"Feast for the eyes" doesn't begin to describe these two bodies of water. Glorious.
I luxuriated in this hammock at Otsego Lake, rocking back and forth, gazing at the leafy sky above me and thinking all sorts of spiritual thoughts. Then two small boys suddenly showed up and began swinging me wildly back and forth, giggling loudly, while I held on for dear life. Such is life as a mother. (I have to admit, I was laughing too — and I remembered that God is found in laughter as much as in silent meditation).
Scott and I slipped out for a lunchtime date, the boys securely in the care of his sister and parents. We decided to try the Sriracha fries with green onion and cilantro, because we love spicy stuff.
Let's just say that if I didn't already believe in God, these would seal the deal. Holy cow, they were good.
Seeing Scott's parents and sister was a treat; we see them so rarely, being on the other side of the country, and I always wish we could change that. But at least we can savor the time we do have. And it was great to see Scott's extended family at the reunion his cousin organized. I met several of his cousins for the first time, and Scott got to see some of them for the first time in about twenty-five years, so it was a blessing for both of us.
Back home in CA, my older son was delighted to finally go get his very own library card. Talk about a rite of passage! As Rita Mae Brown said, "When I got my library card, that's when my life began." It was exciting for him and for book-nerd Mom. I snapped photos like it was prom day.
My mom has sheet music for every occasion.
Two days after returning home, Scott and I celebrated our twelfth anniversary. We had a great dinner at one of our "special occasion restaurants" (otherwise known as a restaurant without a kids' menu) where I ordered quail with truffle risotto (yum). It must have been the quail that got me thinking of this, but I started quizzing him on his knowledge of CA lore, being that he's a native New Yorker and all. He got the state flower right, but he guessed that the CA state motto was "Duuuuude." No wonder I love the guy.
The dotMagis blog is in the middle of its annual month-long celebration of Ignatian spirituality, and they invited me to write about a time when I found God in an unexpected place. My story involves my youngest son, a running leap, the edge of a bunk bed, and an ambulance. You can read it all here. Check out all the other posts, too — there's a wealth of spiritual insight there, and there will be a new one every day of July.
Where have you found God lately?
This entry was posted in In all thINGs, Uncategorized. Bookmark the permalink.


What a fabulous and worthwhile trip you had, Delia! That's really making good use of an extra day off. sorry you hve to put in 10 hr days to get that extra day tho! You really struck it rich with 18 obits! Thanks for telling us about it!


text:
Officers were called to Wince Brook, off Brow Avenue, Alkrington, Greater Manchester, at around 7.10am this morning. The 21-year-old man was pronounced dead at the scene
A 21-year-old man has been found dead at a nature reserve this morning, police have said.
Officers were called to Wince Brook, off Brow Avenue in Middleton, Alkrington, Greater Manchester, at around 7.10am.
It is understood the man's body was found by a dog walker, the Manchester Evening News reports.
Residents in the area said they were awoken by the sound of sirens early this morning.
An ambulance was rushed to the nature reserve, but the man was pronounced dead at the scene.
Greater Manchester Police said enquiries are ongoing but the circumstances around the man's death are not considered to be suspicious.
A GMP spokesperson said: "Police were called around 7.10am today (11 November) to a report of a body having been found close to Brow Avenue, Middleton.
"Emergency services attended and a 21-year-old man was sadly pronounced dead at the scene.
"Enquiries are ongoing but there are not believed to be any suspicious circumstances surrounding his death."
A spokesperson for the North West Ambulance Service said: "We have attended the scene following a call from police at 7:15am, but we haven't taken anyone to hospital."
Wince Brook nature reserve is located near Manchester.
It is a popular spot for dog walkers and also for locals to exercise.
Wallabies wary of ghosts of England past as they seek to avoid relapse


For extra picky squirrels! More pecans, a little extra wheat flour, and a touch of sweetness create a yummy "cookie taste," but still with a low glycemic load.
Ingredients: Pecans, whey protein isolate, wheat protein isolate, oat bran, wheat flour, cracked wheat, whole eggs, tri-calcium phosphate, calcium carbonate, lecithin, baking powder, phylloquinone (vitamin K), selenium, magnesium, vitamin D-3, folic acid, iodine, iron, zinc, copper, molybdenum, manganese, vitamin E, vitamin A (60% beta-carotene), pantothenic acid, niacin, riboflavin, vitamin B-6, thiamin, vitamin B-12, biotin.
My spoiled girl absolutely LOVED these once she actually tried them! So thankful to have been blessed to find Henry's -now my squirrel will get the proper nutrition to keep her healthy!
Our little picky squirrel would only eat the regular blocks if he was super hungry and maybe half at that. He absolutely loves the picky blocks! He sees me get the bag from the freezer and is on my shoulder scrambling to get at them himself! Thank you, Henry's Pets!
My little squirrels love these blocks. They are a great product. They arrive quickly and are very fresh .
My Squirrelly Girl Loves these!
We've ordered the Picky Blocks several times; they are Sugar's favorite "dinner" blocks. Sugar enjoys them so much, she grabs them out of my hand! We also alternate with the Healthy Blocks, but she's less enthusiastic. She started refusing the other "food" blocks; we do not know why. Highly recommended Picky Blocks for the finicky squirrels.
Before I started feeding my squirrel this product, she had gotten to the point of not eating anything I put in her food bowl. She lost weight, which was a scary thing. I found these Picky Blocks online, hoping they would work. I'm happy to report that Ms. Susie LOVES them! I also feed her the Healthy Blocks. She grabs the food from me and immediately chows down. With a mix of some fresh veggies, a few nuts, and the Picky Blocks, my little Susie is a happy, healthy girl.
Was having a problem with my squirrel eating certain foods that were nutritious and finally came across this product to give it a try...so glad I did because he loves them!
My little old man squirrel Loves his Henry's!
My little gray squirrel loves his blocks morning and evening :) Henry's picky blocks keep my little buddy healthy and happy. His winter coat came in so thick and soft, I can't stop snuggling with him!


FORT MEADE, Md. — President Donald Trump is furious after a US Cyber Command offensive cyber operation deleted more than half his Twitter followers, sources confirmed today.
The operation, dubbed Cheeto Fury, targeted the Internet Research Agency, a Russian "troll farm" based in St. Petersburg known for operating fake bot Twitter accounts, among other ways it tries to influence public opinion.
Trump had more than 70 million Twitter followers last week, but the operation brought that number down considerably to roughly 35 million. Of Trump's remaining Twitter followers, 31 million follow only the @realDonaldTrump account, and more than 28 million have not tweeted in the last year.
Duffel Blog freelancer Luke J. Buschlight contributed to this report from Moscow.