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Correlation effect of EGFR and CXCR4 and CCR7 chemokine receptors in predicting breast cancer metastasis and prognosis
Yixin Liu†1,
Ru Ji†1,
Jingyong Li3,
Qiang Gu1,
Xiulan Zhao1,
Tao Sun1,
Jinjing Wang1,
Jiangbo Li1,
Qiuyue Du1 and
Baocun Sun1, 2Email author
© Liu et al; licensee BioMed Central Ltd. 2010
The chemokine receptors CXCR4 and CCR7 play an important role in cancer invasion and metastasis. This study investigated the expression of CXCR4, CCR7, CXCL12, CCL21, and EGFR to illustrate the role of these biomarkers in breast cancer metastasis and prognosis.
The CXCR4, CCR7, CXCL12, CCL21, and EGFR biomarkers were analyzed along with ER, PR, and HER-2/neu in breast cancer tissue microarray (TMA) specimens, including 200 primary breast cancer specimens by immunohistochemistry. Corresponding lymph nodes from the same patients were also examined using the same method.
Together with their CXCL12 and CCL21 ligands, CXCR4 and CCR7 were significantly highly expressed in tumor cells with lymph node (LN) metastasis. Similarly, EGFR was expressed highly in tumors with LN metastasis. The ligands were especially expressed in metastatic tumors than in primary tumors from the same patients. Moreover, the expression of both CXCR4 accompanied by CCR7 and CXCL12 accompanied by CCL21 were up-regulated. Kaplan-Meier survival analysis revealed that patients exhibiting high CXCR4, CCR7, and EGFR expression experienced a shorter survival period compared with those with low expression.
The expression of CXCR4, CCR7, and EGFR may be associated with LN metastasis. Moreover, the expression of these receptors can serve as an indicator of undesirable prognosis in patients with breast cancer.
Epidermal Growth Factor Receptor Expression
CXCR4 Expression
Breast Cancer Metastasis
Breast cancer ranks among the most common malignant tumors afflicting women worldwide. Despite decreased mortality rates resulting from combined therapy, breast cancer remains a leading cause of cancer death in women. Particularly in the last two decades, incidence and mortality rates of breast cancer have climbed sharply in China, thus attracting increased attention from researchers.
Metastasis is one characteristic of malignant tumors which determines the course of therapy and cancer prognosis. It is a multifactorial, nonrandom, and sequential process with an organ-selective characteristic. In essence, axillary lymph node metastasis is the most frequently occurring metastatic disease; it can be seen as a surrogate for distant metastasis and long-term survival [1].
Although several molecules are involved in breast cancer metastasis, precise mechanism of tumor cell migration to specific organs remains to be established [2]. Previously, the "seed and soil" theory was employed to explain directional metastasis, considering that certain metastasis organs possess the congenial environment of the primary organ [3]. More recently, a "chemokine-receptor" model has been proposed to explain the homing of tumor cells to specific organs [4]. Chemokines belong to a super-family of small, cytokine-like proteins that induce cytoskeletal rearrangement and adhesion to endothelial and directional migration through their interaction with G-protein-coupled receptors [2, 5].
Among the chemokines, the most interesting chemokine-receptor pair is the CXC chemokine receptor-4 (CXCR4) and its lone ligand, CXC chemokine ligand-12 (CXCL12). Muller demonstrated that CXCR4 is consistently expressed in human breast cancer cells, malignant breast tumor and metastasis tumors, while its ligand CXCL12 is preferentially expressed in the lungs, liver, bone marrow, and lymph nodes [2]. Thus, it can be deduced that the CXCL12-CXCR4 axis may be associated with the metastasis of breast cancer cells to the lungs, liver, bone, and lymph nodes. Unlike CXCL12, however, CC chemokine ligand-21 (CCL21) - the ligand for CC chemokine receptor-7 (CCR7) - is highly expressed in the lymph nodes of breast cancer patients [5]. Thus, the CCR7-CCL21 axis can be said to assume an important role in lymph node metastasis [6]. In this study, the expression of both CXCR4 and CCR7 is combined to evaluate their contribution in the lymph node metastasis of breast cancer. The importance of growth factors such as epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor2 (HER-2/neu) has been established in the prognosis of breast cancer. Recently, several studies have revealed the crosstalk between CXCR4 and EGFR or HER-2/neu through transactivation by the CXCL12-CXCR4 axis.
This study aims to verify the significance of CXCR4, CCR7 and their CXCL12 and CCL21 ligands, together with EGFR in the evaluation of metastasis and the prognosis of breast cancer.
Patient selection and clinical data
The study group was composed of 200 specimens selected from 284 cases (84 cases were excluded owing to the absence of follow-up status) of female primary invasive duct breast cancer cases diagnosed between January 1997 and December 2004 at the General Hospital of Tianjin Medical University. Patients' records were retrieved and clinical data, histopathological record, and treatment information were all reviewed. All patients had not been subjected to chemotherapy and radiotherapy prior to surgical resection but had received chemotherapy following surgical operation. Follow-up information from all the patients were obtained by the authors themselves in August 2009 through visits or telephone interviews with either the patients or their relatives. Mean follow-up time was 88 months, ranging from 5 to 150 months. Formalin-fixed paraffin-embedded tumor materials and their lymph node tissues were acquired from the Department of Pathology of Tianjin Medical University's General Hospital. Tumor diameter, pathologic stage, and nodal status were selected from the primary pathology reports. All slides were reviewed by two pathologists to define histological types and grades.
Construction of tissue microarray
Tissue microarray (TMA) blocks were constructed from formalin-fixed, paraffin-embedded breast cancer samples stored at the Department of Pathology of Tianjin General Hospital. These TMA blocks were composed of 200 paired samples (primary tumors and corresponding lymph nodes, either metastatic or non-metastatic). Haematoxylin and eosin stained slides were reviewed to confirm the diagnosis of invasive breast cancer; afterwards, two representative tumor regions were selected and marked on the donor blocks.
Tumor TMA blocks were created by punching a cylinder using a hollow needle with a diameter of 2 mm; the blocks were obtained from the two selected areas of each donor block before being inserted into an empty paraffin block. Subsequently, these blocks were cut into 4 μm thick slides and prepared for immunohistochemical (IHC) analysis.
Immunohistochemical analysis
Using IHC staining, the expression of different proteins in human breast cancer was verified. In this process, sections were deparaffinaged in xylene prior to rehydration using gradient alcohol. Endogenous peroxydase activity was blocked by 3% hydrogen peroxide in 50% methanol for 20 minutes. For antigen retrieval, sections were treated with citrate buffer saline (pH 6.0) for 15 minutes at 95°C in a microwave oven. After blocking with 10% normal goat serum for 30 minutes at room temperature, sections were incubated with primary antibodies for another 30 minutes at room temperature. The sections were subsequently incubated for 16 hours at 4°C. Primary antibodies and dilution were as follows: rabbit polyclonal anti-CXCR4 (Abcam, dilution 1:100); rabbit polyclonal anti-CCR7 (Abcam, dilution 1:100); rabbit polyclonal anti-CXCL12 (Abcam, dilution1:100); goat polyclonal anti-CCL21 (Santa Cruz Biotechnology, dilution 1:50); rabbit polyclonal anti-EGFR (Santa Cruz Biotechnology, dilution 1:100); mouse monoclonal anti-ER (Zhongshan; ready-to-use); rabbit polyclonal anti-PR (Santa Cruz Biotechnology, dilution 1:100); and mouse monoclonal anti-HER2 (Zhongshan; ready-to-use).
Following incubation, sections were lavaged with phosphate buffered solution (PBS) and incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG, goat anti-mouse IgG, or rabbit anti-goat IgG for 40 min at room temperature. Staining was performed using 3,3'-diaminobenzidine (DAB). Sections were counterstained with haematoxylin followed by dehydration and mounting. Negative controls were prepared using PBS in lieu of the first antibody.
Scoring of immunostaining
Sections were read by two separate pathologists blinded to patients' clinical pathology parameters. Both intensity and percentage of positive cells were considered. Five microscopy fields were reviewed in each core with 400× magnification, after which positive cells of 100 tumor cells in each field were counted. In staining for CXCR4, CCR7, CXCL12, CCL21 and EGFR, tumor cells with brown cytoplasm and/or nucleus or membrane were considered positive and then scored based on four classes: none (0); weak brown (1+); moderate brown (2+); and strong brown (3+). Percentage of stained tumor cells was categorized into five classes: 0 for negative cells, 1 for 1-25%; 2 for 25-50%; 3 for 50-75%; and 4 for >75%. Multiplication (staining index) of intensity and percentage scores was utilized to determine the result.
A staining index of ≥6 was defined as high expression, while <6 was defined as low expression [7]. On the another hand, HER2/neu was evaluated as positive when over 10% of tumor cells exhibited stained consecutive membranes. Unified standards were employed when evaluating estrogen receptors (ERs) and Progesterone receptors (PRs) that exceeded 10% of tumor cells, as shown in the stained nucleus.
Analyses were performed using the SPSS 17.0 software package (Chicago, IL, USA). The relation between CXCR4, CCR7, EGFR, and clinicopathologic characteristics were tested via Pearson χ2 analysis. The same method was employed to test associations between these biomarkers and biologic-prognostic characteristics, such as ER, PR, and HER-2/neu expression. Correlations between two variables were evaluated by Spearman's rank correlation test. P-values < 0.05 were deemed statistically significant. Overall survival (OS) was estimated through the Kaplan-Meier method and was compared between groups through the log-rank test.
Characteristics of patients and expression of biomarkers in primary tumors
Patient and primary tumor characteristics are presented in Table 1. Samples included 200 patients, among which 100 developed lymph node metastasis while 100 did not. Median age was determined at 51 years (37-74). Thirty-nine patients (19.5%) were diagnosed with stage I cancer, 138 (69%) with stage II, 20 (10%) with stage III, and three (1.5%) with stage IV.
Correlation between biomarkers and primary tumor characteristics
CXCR4 cytoplasmic expression
CXCR4 nuclear expression
CCR7 expression
EGFR expression
tumor size
D≤2
2<D≤5
D>5
.030*
<.001**
N≤3
3<N≤10
N>10
HER2/neu
.005**
D represents the diameter of tumor, LN represents lymph node status, N represents the amoumt of lymph node excised, grade means the histological grade, and stage means the clinical stage. *P < 0.05, **P < 0.01
In IHC staining, 77% of tumor cells were CXCR4 positive in the cytoplasm, including high and low CXCR4 expression (Figure 1A2). Meanwhile, 73% were positive in the nucleus (Figure 1A2). The amounts of CCR7 (Figure 1B2) and EGFR (Figure 1E2) were detected in 82% and 66% of tumor cells, respectively, in the cytoplasm and/or membrane. Furthermore, 50% of ER, 49.5% of PR, and 23.5% of HER-2/neu were observed to be positive.
IHC staining for biomarkers. IHC staining for CXCR4, CXCL12, CCR7, CCL21 and EGFR. PT pertains to primary tumor, while LNMT stands for lymph node metastasis tumor. Rows correspond to the designated chemokine or receptor. The first column represents staining of negative expression in primary breast cancer with the indicated antibody. The second column indicates positive expression in primary breast cancer, and the third column shows positive expression in lymph node metastasis cancer. Both PT and LNMT columns in each row are obtained from the same patient while the negative column is not. In the CXCR4 row, A2 and A3 exhibit high expression in both cytoplasm and nucleus. CCR7, CXCL12, and CCL21 all exhibit positive reaction in the cytoplasm. In the EGFR row, E2 and E3 indicate that EGFR is expressed mainly in the membrane. However, a number of tumor cells appear to be positive in the cytoplasm as well (Panels A-E, ×200).
Association of CXCR4, CCR7, and EGFR with lymph node metastasis
The immunoreactivity of CXCR4 was observed in the cytoplasm and/or nucleus of tumor cells. Cytoplasmic reactivity of CXCR4 correlated positively with lymph node metastasis of breast cancer (P < 0.001), but not with the amount of involved lymph nodes. Nuclear reactivity was not observed to be correlated with any pathologic parameters. Meanwhile, CCR7 was positively expressed in the cytoplasm, and the activity was significantly correlated with lymph node metastasis (P < 0.001). Similarly, associations among the lymph node status, histological grade, and EGFR expression were observed in this study (Table 1).
To verify the important effect of CXCR4 and CCR7 in metastasis, CXCR4, CCR7, and EGFR expression in primary breast cancer were compared with that in lymph node metastasis tumor. It was observed that CXCR4 and CCR7 expression in metastasis tumor was even higher, although no significant distinction was evident. More importantly, their respective ligands, CXCL12 and CCL21, exhibited significant differences in expression between primary tumor and lymph node metastasis tumor (P = 0.016 and P = 0.004; Table 2). Distinction between chemokines and their receptors with regard to distribution may be associated with the mechanism of metastasis; specifically, chemokines attract their receptors to certain sites along the chemokine concentration gradient.
Differences of biomarkers between primary tumor and lymph node metastasis tumor
cytoplasmic CXCR4
CXCL12
CCL21
EGFR
PT means primary tumor, MT means lymph node metastasis tumor. The differences of the biomarker between primary tumors and metastasis tumors were tested by pearson χ2 analysis. *P < 0.05, **P < 0.01
Correlation between CXCR4, CCR7, EGFR and HER-2/neu
Although neither ER nor PR positivity was associated with degree of the biomarkers, HER2 over-expression was correlated with CXCR4 cytoplasmic positivity (p = 0.039; Table 1). As indicated by reports, the expression rate of HER2/nu in breast cancer is approximately 25%. In the results of this study, the expression of HER2 was nearly 20%, and among CXCR4 cytoplasmic positive patients, approximately 40% were associated with HER2 expression. In summary, tumors positive for CXCR4 cytoplasmic staining are more likely to be positive for HER2 over-expression.
As an independent prognostic factor for breast cancer patients, EGFR is associated with a number of pathological characteristics of breast cancer. According to the results, EGFR expression is correlated with lymph node metastasis and histological grade (Table 1). Interestingly, during analysis, it was discovered that close to 70% of patients with high EGFR expression were CXCR4 and CCR7 positive as well. Spearmam's rank correlation analysis revealed that EGFR expression was significantly associated with CXCR4 cytoplasmic positivity and high CCR7 expression (P < 0.01; Table 3).
Correlation of CXCR4, CCR7 and EGFR
CXCR4 cytoplasmic and EGFR
<0.001**
CXCR4 nuclear and EGFR
CXCR4 cytoplasmic and CCR7
CXCR4 nuclear and CCR7
0.008**
CCR7 and EGFR
The correlation between every two biomarkers was tested by Spearman's rank correlation test. *P < 0.05, **P < 0.01
Concordance of CXCR4, CXCL12, CCR7, and CCL21 expression
After performing IHC staining for the two CXCL12 and CCL21 chemokines, it was revealed that these were correlated with one another (P = 0.017, Table 4), indicating a tendency towards co-expression of these molecules in tumors. Hence, the expression of their receptors, CXCR4 and CCR7 was likely to be tightly linked (P = .008; Table 4). No significant association was present between the expression of CXCR4 and CXCL12, nor between CCR7 and its chemokine ligand CCL21 (Table 4).
Correlation of CXCR4, CCR7 and their ligands CXCL12, CCL21
CXCR4 cytoplasmic and CXCL12
CCR7 and CCL21
CXCL12 and CCL21
Correlation was tested by Spearman's rank correlation test. *P < 0.05
CXCR4, CCR7, and EGFR demonstrate poor prognosis by survival analysis
Follow-up investigation revealed that the median survival time was 88 months (ranging from 5-150 months), within which 45 patients (22.5%) died because of breast cancer including 28 (28%) in the tumor with metastasis group and 17 (17%) in the non-metastasis group. Kaplan-Meier analysis revealed that patients suffering from high levels of CXCR4 expression- either in the cytoplasm or in the nucleus -had significantly lower OS compared with those with low CXCR4 expression (P = 0.011, Figure 2; P = 0.003, Figure 3). Similarly, high levels of CCR7 and EGFR expression revealed poor prognosis (P = 0.044, Figure 4; P = 0.007, Figure 5).
Overall survival analysis for CXCR4 cytoplasmic expression. Kaplan-Meier curves for overall survival (OS) in 110 patients with high expression of CXCR4 and 90 patients with low expression of CXCR4 in cytoplasm. Survival time sharply decreased in patients with high CXCR4 cytoplasmic expression, especially in the first five years, Meanwhile, survival of patients with low CXCR4 expression was merely moderately affected (P = 0.011).
Overall survival analysis for CXCR4 nuclear expression. Kaplan-Meier curves for overall survival (OS) in 113 patients With high CXCR4 expression and 87 patients with low CXCR4 expression in the nucleus. Survival time sharply decreased in patients with high CXCR4 nuclear expression, especially in the first five years, when significantly compared with those exhibiting low expression (P = 0.003).
Overall survival analysis for CCR7 expression. Kaplan-Meier curves for overall survival (OS) in 111 patients with high CCR7 expression and 89 patients with low CCR7 expression in the cytoplasm. The difference between these two groups is not highly significant as determined by the log-rank test (P = 0.044). However, it can be observed from the curve that in the first five years, survival rate sharply decreased in patients with high CCR7 expression in the cytoplasm, while hardly any patient in the low expression group died during the first five years.
Overall survival analysis for EGFR expression. Kaplan-Meier curves for overall survival (OS) in 88 patient with high EGFR expression and 112 patients with low EGFR expression in the membrane and cytoplasm. Survival rate of patients with high EGFR expression was significantly low compared with those exhibiting low expression (P = 0.007).
Recently, reports have demonstrated that chemokines and their receptors play critical roles in the development of cancer, including tumor cell growth, migration, and angiogenesis. Further, they influence the infiltration of immune cells in a tumor [8, 9]. The mechanism of chemokines in malignant tumor metastasis may be reflected by the production of chemokine receptors by tumor cells, which respond to their homologous ligands (produced by certain organs) and migrate along the chemokine gradients to trigger specific organ metastasis [10]. Out of all the known chemokine receptors, breast cancer cells specifically express active CXCR4 and CCR7, the ligands of which are HCXCL12 and CCL21, respectively [2].
This study investigated a series of matched primary and lymph node metastasis breast cancer tumors to demonstrate whether the expression of the CXCR4 and CCR7 chemokine receptors, along with expression of EGFR, predicts increased risk of metastasis and mortality. Present data are consistent with those in previous reports describing a positive correlation between CXCR4 expression and lymph node metastasis in cases of non-small-cell lung cancer (NSCLC), nasopharyngeal cancer, colorectal cancer, and esophageal cancer [11–14]. Positive correlation has likewise been reported between CCR7 expression and lymph node metastasis in cases of NSCLC, breast, gastric, colorectal, esophageal, and thyroid cancer [15–20].
It has been demonstrated that the CXCR4/CXCL12 axis likewise induces chemotaxis and breast cancer cell migration. Since Muller reported that CXCR4-CXCL12 interaction governed the pattern of breast cancer metastasis in a mouse model, subsequent studies have been conducted in different tumors [2]. One study determined that the CXCR4 expression pattern correlated significantly with the degree of lymph node metastasis by investigating CXCR4 expression in 79 cases of invasive duct cancer (IDC) [21]. Su examined 85 cases of early breast carcinoma and learned that high cytoplasmic expression of CXCR4 is associated with axillary nodal metastasis [22]. In the prent study, CXCR4 was found to be present in both cytoplasm and nucleus of tumor cells, and cytoplasmic expression was associated with lymph node metastasis. This result is similar to that of certain studies [22–25], but is contrary to a handful of reports [26]. Further, CXC chemokine 12 (CXCL12, likewise known as stromal cell derived factor-1α, or SDF-1α) is expressed in the liver, lungs, brain, bone, and lymph nodes. on the other hand, CXCR4 is a membrane-bound G-protein-coupled receptor which, together with its ligand CXCL12, mediates inflammatory and tumor cell migration [27]. One study has also observed CXCR4 localization at the cytoplasm in leukocyte cell lines with enforced CXCR4 expression and CXCL12-induced polarization of CXCR4 to the edge of migrating leukocyte cells [25]. Hence, with regard to the effect of CXCL12, CXCR4 reactivity in the cytoplasm may reflect receptor internalization. This may be viewed as an activation state of CXCR4. Through immunohistochemistry, CXCL12 protein in the cytoplasm of tumor cells was located as well, and CXCL12 expression was observed to be higher in lymph node metastasis tumors than in primary tumors. This distinction in expression sites between chemokines and their receptors illustrates that CXCL12 attracts CXCR4 to certain metastatic sites along the concentration gradient.
Although nuclear CXCR4 is expressed in cancer cells, its function remains unclear. Spano demonstrated that nuclear CXCR4 expression represents a better outcome in patients afflicted with non-small-cell lung cancer [28]. However, in the present study, after over 10 years of follow-up observation conducted among 200 breast cancer patients, it was noted that high expression of both cytoplasmic and nucleus CXCR4 often indicated worse prognosis. Different localization patterns of chemokine receptors-whether nuclear or cytoplasmic-may have different levels of biological significance in cancer cells.
Similarly, the interaction between CCL21 and its CCR7 receptor plays a crucial role in lymphocytes homing to secondary lymphoid organs through lymphatic vessels. A study indicates that the hindrance of T cells homing to secondary lymphoid organs occurs because of the loss of CCL21 or the deletion of the CCR7 gene [29]. Hence, it is likely that the mechanism of CCL21 mediating migration of tumor cells to lymph nodes from primary site arising from its attraction to CCR7, which is highly expressed by primary tumors, is similar to the mechanism of the lymphocytes' homing effect. Results of this study revealed that 70% of primary breast cancer tissues and 77% of metastasis cancer cells in lymph nodes expressed CCR7. Further, there was a significant correlation between CCR7 expression and lymph node metastasis (p < 0.001); CCL21 was especially highly expressed in lymph nodes metastasis tumor cells (68%), which was not the case in primary tumor cells (P = 0.004).
Survival analysis revealed patients with highly expressed CCR7 are subject to a more undesirable prognosis compared with those who expressed low CCR7. Findings of this study coincide with those of other studies [7]. In view of all the evidences, there is reason to believe that the CCR7-CCL21 axis is a crucial factor in tumor lymph node metastasis. Moreover, as staining for CXCL12 and CCL21 (or CXCR4 and CCR7) was tightly linked in the group of primary tumors and lymph node metastasis tumors in this study, it is likely that a shared mechanism may account for variations in expression levels of both molecules in breast cancer. Coinciding with previous studies, it was demonstrated that levels of combined CCR7 and CXCR4 expression significantly correlated with lymph node metastatic status[16, 17, 30].
Recent studies and analyses conducted in the present study clearly indicate that EGFR expression serves as the strong prognostic factor in invasive breast cancer [23, 31, 32]. In this study, it was observed that patients with high EGFR expression are more prone to developing metastasis and possessing high grades of tumor, which are both important prognostic factors for breast cancer patients. Through survival analysis, it has been discovered that patients who highly express EGFR are subject to poor prognoses compared with those with low EGFR expression. Recent reports further suggest that CXCR4 expression can be up-regulated by HER-2/neu, which is required for HER2-mediated invasion in vitro and lung metastases in vivo [33]. Moreover, the result of the correlation between CXCR4, CCR7, EGFR, and HER-2/neu illustrates that the expression of chemokine receptors (CXCR4 and CCR7) is tightly associated with growth factors (EGFR and HER-2/neu). Based on this finding, it may be inferred that regulating growth factors may influence the expression of chemokine receptors, which may be helpful in identifying new pathways in breast cancer therapy.
This study was based on a small group of patients. However, it examined corresponding lymph nodes of each patient, and this has not been reported by other scholars to date. Although immunochemistry detection of the biomarkers may have certain limitations, it is a simple and widely utilized technique which can be carried out on routine paraffin-embedded tissues. By contrast, majority of new biological methods require specialized platforms and expertise that are considered impractical in routine pathological diagnosis.
By examining the expression of chemokines and their receptors in both primary tumors and corresponding lymph node metastasis tumors, data indicate that chemokines and their receptors are differentially expressed in the primary and metastatic sites of breast cancer. Results reveal the significant association of CXCR4, CCR7, and EGFR with metastasis and poor prognosis. Further, the correlation between chemokine receptors and growth factors may provide a new method of understanding breast cancer metastasis and therapy, which are worthy of further study.
Yixin Liu, Ru Ji contributed equally to this work.
The work was supported by grants from the Tianjin Natural Science Foundation (Nos.06YFJMJC08000 and 09ZCZDSF04400), as well as a grant from a key project of the Natural Science Foundation of China (No.30830049). Materials were obtained from the Department of Pathology of Tianjin Medical University's General Hospital.
Before submission, all authors read and approved the final manuscript. Among the authors, LYX designed the study, while JR collected the materials, performed all experiments, and drafted the manuscript. LJY conducted the statistical analysis and GQ accomplished construction of tissue microarray blocks. ZXL participated in the instruction of the experiment, while ST revised the manuscript critically to ensure important intellectual content. WJJ and LYX read and reviewed the sections, while, LJB and DQY performed follow-up observations on all patients. SBC provided the study concept and participated in its design and coordination.
Department of Pathology, Tianjin Medical University, Tianjin, China
Department of Pathology, Tianjin Cancer Hospital, Tianjin Medical University, Tianjin, China
The College of Public Health, Tianjin Medical University, Tianjin, China
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There are huge numbers of differences between us and the writers and first hearers of Scripture. But most of them don't matter most of the time. It's the ones that do matter that really damage our reading.
This video also deals with how our expectations (preunderstandings: what we expect the passage to be saying) can lead is astray.


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Home › All Patterns › Uhura Crochet Graphghan Pattern
Uhura Crochet Graphghan Pattern
From The Graphalo - Fandom and pop culture crochet and knitting patterns perfect for everyone from absolute beginners to experts.
Nyota Uhura is a character in the Star Trek franchise. In the original TV series, the character was portrayed by Nichelle Nichols who later reprised the role for the first six Star Trek feature films. Since 2009, Uhura has been portrayed by actress Zoe Saldana.
Whoopi Goldberg, who later played Guinan on Star Trek: The Next Generation, described Uhura as a role model for her, recalling that she told her family, "I just saw a black woman on television; and she ain't no maid!" NASA later employed Nichols in a campaign to encourage women and African-Americans to join the service. NASA Astronaut Group 8 yielded the first recruits composed of women and ethnic minorities—three were black (Guion Bluford, Ronald McNair and Dr. Frederick D. Gregory). Dr. Mae Jemison, the first black woman to fly aboard the Space Shuttle, cited Star Trek as an influence in her decision to join. Jemison herself had a minor role on an episode of The Next Generation called "Second Chances", playing a transporter operator named Lieutenant Palmer. Jemison was the first of several real-life astronauts to appear on Star Trek. When Nichols considered quitting the show in the late 1960s, she was told someone wanted to meet her, claiming to be her biggest fan. The fan was Martin Luther King Jr., who convinced her not to leave the show because "You cannot do that. ... For the first time, we are being seen the world over as we should be seen." Nichols decided to stay on the show, explaining her reasons to Roddenberry, who replied "finally, someone gets it."
Pattern by @TheGraphalo
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redpajama_set_name: RedPajamaCommonCrawl


... with the Meson build system on Linux, and ~15x faster on Windows.
Last year, Jussi Pakkanen (the maintainer of Meson) gave a talk about improving the way GNOME apps are built by replacing Autotools with the Meson build system.
This year, I'll be talking about how we at Centricular worked along exactly those lines and ported GStreamer, GLib, and its core dependencies from Autotools to Meson. Besides the improvements in speed, using Meson provided numerous advantages over Autotools such as cleaner syntax, better cross-platform support, maintainable code, and much more.
I will be talking about our porting journey and make the case that Meson is indeed ready for replacing Autotools in every case we've seen so far. The next step is convincing more projects to try out Meson.


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Mary Anne Evans was born in England, the third daughter of Robert and Christiana. Two of her siblings were already teenagers when she was born, from a previous marriage of her fathers. Her other siblings were born of both her parents as well, a sister and a brother.
She was considered an intelligent child, and, her father, being employed as the manager of a large Estate entitled her to use it's library, which helped her in her early education. She was raised Anglican, but also saw a growing number of religious dissenters in the area of the Midlands where she was raised.
She attended boarding school until her mother passed away, and she returned home to fulfull the housekeeping role. Her older brother, Isaac eventually married, and took over the family home, so she and her father moved to Coventry. There, she met a couple named Charles and Cara Bray, whom were rich from ribbon manufacturing. They were very free thinking and philanthropic, and their home frequently housed people who enjoyed debate, and radical views. She met many people, like Ralph Waldo Emerson, while she was present in their home. Through the myriad of people, and debate, Mary Anne lost her faith, and her father threatened to toss her out. He never did, because she respectfully attended church, and kept his house for him, until his death. Less than a week after his death, she went to Switzerland with the Brays, and decided to stay in Geneva, by herself. She eventually returned to England, living in London, with the intent to be a writer. She became instrumental in helping to edit a left wing journal called "The Westminster Review", for which she also wrote many essays and reviews.
This being the 1800's (in case you were wondering), it was scandalous to many to have an unmarried woman interacting with the predominantly male society of London. She was clearly strong minded, though, very sensitive, often depressed, and had much self doubt. She was not considered attractive, by any means, and frequently fell in love with those who did not reciprocate. She eventually met a man who loved her back, George Henry Lewes, a philosopher, and critic. George was married, but he and his wife had an "open" marriage. George and Mary Anne, (now Marian) chose to live together. (Lest you think George's wife was neglected, she had three children with George, then, MORE children with other men.) He was unable to divorce his wife because he was named as father for at least one of her offspring with another man, because this was considered 'complicit in adultery'.
She and George went to Germany, for research, and considered the trip a "honeymoon". They styled themselves as married, and she called herself by his last name. Their open ness with their affair added to her scandalous reputation. They lived apart from literary society, both shunning, and being shunned equally. She continued to contribute to the Westminster Review, but resolved to be a novelist. Not wanting to be included into the cache of women who were currently writing, whom she considered silly, and trivial, she chose a pen name, George Eliot.
Her work was an instant success, and as she was using a pen name, people were speculating a great deal to her identity. Pretenders even came forward to claim her work. She eventually had to step forward, and claim it herself. Though her readers were shocked, they did not stop reading her novels. It took a very long time for society to accept her, and her husband, though, they did. She was eventually granted an audience with the daughter of Queen Victoria, who was an avid reader of her novels.
She continued to write, prolifically, for fifteen years. Her husband died, and she spent two years editing HIS final works. During this time, she met a man, twenty years junior to her, John Walter Cross, an American banker. She married him (legally, this time), shocking every one. They honeymooned in Venice. Sadly, he wasn't a stable person, and either jumped, or fell from their balcony into the Grand Canal. He did survive. She, however, fell ill, and, compromised already by kidney disease, she died at the age of sixty one.
Died at the age of... ?!!! GAH!
Anyway, loved it. I've never delved into this one. :)
I had no idea .. you always open my eyes with these posts!
Queenie Tuesday
Monday Morning Mush
zzzzzzzzzzzzzzzz
The Queens Tuesday....
Happy Birthday, Dad!
In which I am going to rant, a little
Bloggingham Love Shack (aka The Romance Meme)1. Wh...


text:
Edging in close to the release of his next EP A Sign of Things to Come, Tom Perrett AKA Advance is more than excited to open up about his next futurepop gem. Blending sci-fi elements with social commentary as well as creating club beats that will make any person on the dance floor move their feet, Advance is not ashamed about his latest production. In fact, he goes on to say it's even stronger than his debut album Deus Ex Machina. And if all that isn't enough to get your appetite whet, perhaps an unreleased track from A Sign of Things to Come will. Stream this banger right below and let it be the soundtrack to Advance's words of wisdom below!
Hey there Tom! Lovely to finally chat with you after gobbling up your releases time and time again. How have things been with advance these days?
Tom: Hey, thanks for having me. Advance has been going well. I've been working tirelessly to get the new album finished which now has a title and I think all of the tracks are written too, so we will be looking at getting that done probably early next year. A Sign of Things to Come is just around the corner which the label and I had decided to put out after us deciding that getting the album done this year was highly unlikely. All the same it's four tracks that I'm very proud of - it feels like a step up in terms of quality and the extra remixes from Biomechanimal and Nature of Wires on the digital version are fantastic.
I'd like to take a trip down history lane. You have an amazing EBM/Futurepop crossover going on with Advance. When was it that you were first exposed to the genre? Was there any one band you remember listening to and thinking, "I need to make music like that."
Tom: Three bands actually. I fell in love with the sound of VNV Nation, Assemblage 23 and Neuroticfish, and that introduced me to EBM / Futurepop initially. It all happened more or less by accident, Advance was never planned so it's a purely organic project in a sense. I started recording different things on a DAW I was using at the time, created an electronic piece and people seemed to like it. From there I decided I would play about with different things and make a couple of rough demos and again certain people seemed to like it. It really all hit when Barry Douglas (COG Promotions in Glasgow) offered me the local support slot for Assemblage 23 and it was then I started taking it seriously. Funnily enough I didn't even have more than three songs written, songs like 'Dead Technology' had to be written to make sure we could play live. At the time I had three songs; 'Fractured Existence' (the first Advance song ever), 'Enter the Wastelands' and 'When We Return'. I wrote 'Apathy' and 'Dead Technology' in a few days.
While Advance's music is all you, your live line-up also consists of Kimberly Copland on synths. Who is she, how did you meet, and why did you choose her for synths during live performances?
Tom: Kim has always been there - she is my other half for those who don't know. She joined as a live member in 2014 and has been involved since then. With the release of the new EP and after the great working relationship we have I decided to include her as a full member. This means in future she will also help with the writing process and I'm fairly certain that people will love what she has to offer. We have been in a relationship for seven plus years actually and we met through a mutual love of this style of music, so it's kinda poetic in a way.
When was it that you first created Advance? And why choose that particular name? Did you ever think of calling your band anything else?
Tom: If I did think of another title at the time, I'm not sure what it would have been, I settled with Advance fairly quick. I would love to be able to give a great reason for the name but honestly I'm not even sure how I settled on the name; it's a name which conjures up different connotations for different people and I like that quite a lot it leaves it open to interpretation.
Looking at your Discogs page, I've only found your label debut Deus Ex Machina on there. Do you have any previous releases, singles, or demos floating around on the internet? If so, do you plan on ever re-releasing that material to the public?
Tom: There is a demo somewhere with three tracks called 'Dead Technology' but I can't see me ever re-releasing anything from back then. Because of how I work I lose a lot of ideas through switching DAW or PC or whatever else that can effect workflow, and I think I've binned more projects than I've saved.
So, let's talk about your latest EP that's soon to come out A Sign of Things To Come. The cover art easily showcases a Blade Runner-esque environment, and it has already been stated that the album has social commentary in it. What messages did you put in A Sign of Things To Come and how did you blend it with a sci-fi theme?
Tom: I'm interested in people's ignorance to their surroundings, A Sign of Things to Come is about greed, plain and simple, and is filled with lyrics like, "We are held in place, like parts of a machine/ we are all just human batteries to feed the elite". It follows this path of how easily people are bought, how people have no real conviction anymore and how control of your own life is an illusion.
How did you approach writing A Sign Of Things To Come musically? What did you do differently to make this EP stand out from Deus Ex Machina?
Tom: I suppose the biggest difference was time, and taking time to get things just right. After DXM I spoke to Adrian the label head and we discussed time periods for the next release. The AnalogueTrash guys are great - they give you time to do right by the project and make sure what you are releasing is worthwhile. I have a great working relationship with AT and one which will probably continue for a long time.
I think I may have touched this above, but how did the lyrics differ in substance from your previous work? Did you feel as if anything presented in this album overlapped in the previous?
Tom: Interesting point, and I would say it's another take on a hopeless society, but this time more contemporary. DXM is all apocalyptic and almost set in the future I guess, and this one is more grounded in reality, or at least our current reality.
I always like asking this questions to musicians, but, when you finally heard A Sign of Thing to Come in its full, mastered form, what did you think of it? Did you find perfect? Did you see any errors? What is your
critique on the album?
Tom: Mostly I heard a lot of positive improvements from the first album. The material is stronger in my opinion and it's more in line with where I want Advance to be. I'm not saying that all the albums will be political because they won't, it will meet whatever subject I think suits the material best. Nothing is ever perfect though. The curse of making music is that you are always improving and by the time you release something you have already improved since that point as well so things are always slightly behind that curve. Although, in saying that I think that it's more important how other people view it, we have seen in the past plenty of times where a band thinks something is their best work and it alienates the people who loved the band so it fails ultimately. It's a tricky one really.
We already gave an early review to the album and gave it a solid 8 out of 10. However, how has reception been elsewhere? Have critics/fans been favoring this EP?
Tom: It's early days but all the feedback so far is really good and it seems to be well received. Ask me that again in a couple of months, ha ha!
And right now what are your future plans? Do you have any live shows/gigs scheduled? If so, where? I also understand you have a full length album scheduled for 2017. Could you give us any information on that?
Tom: This is the first time in ages where I've had no gigs planned, not even local ones. I'm focusing on getting the album done really. If the right offer comes in, I'll have a look and see what can be done but I don't just accept gigs for the sake of gigging if I can help it. That said, I'm hoping we'll play our first show outside the UK this year. Information about the album ... Short answer ... no but the long answer is it's going well. I think all the songs are nearing completion but I would like to take some time to go over them very carefully, cut out bits of filler and have an album which is something I will be happy to release. I have a title but I'm going to keep that under my hat for now.
Lastly, I'd like to thank you for your time and wish you the best! You may use the space below to mention anything I may have failed to cover. Cheers!
Tom: It's my pleasure. I hope all of you enjoy A Sign of Things to Come and I would like to thank everyone who supports the project I am immensely grateful for it. Hopefully I'll see some of you on the road soon.
A Sign of Things to Come is available for pre-order HERE via AnalogueTrash.
Advance interview
Steven Gullotta https://www.brutalresonance.com/interview/interview-june-2016/ June 1, 2016 June 1, 2016
EBM, Futurepop
# advance
# ebm
# futurepop
# glasgow
# scotland
# a-sign-of-things-to-come
Gioumourtzina - 'Blakk Metall'
Blast Radius Releases New Video Single "When It Works"
What's the point of living if it's to copy and repeat over and over again what we've been taught ? This is bullshit ! We're in this world to become ourselves and to free our mind from our education and our own self-centred behaviour and blindness, not to train ourselves to become another clone...
Krystal System, Mar 06 2014
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SEATTLE — Morning thunderstorms are slamming into the Washington coast with dangerous lightning while heavy rain dumps inland — creating problems for commuters.
Today: Heavy rain showers are tracking through with a building southerly wind. Wind gusts for Everett to Seattle to Bremerton and Olympia will be 30 to 45mph and a Wind Advisory has been issued until 6pm tonight for most of the inland cities. Stronger wind gusts up to 60mph have kept a high wind warning in place for western Whatcom, Skagit, Island counties.
Tonight: Friday Night the weather keeps some rain showers, wind only breezy at times.
Rain will increase again later Saturday morning and this will be a round which dumps another 1-3″ of rain over central Puget Sound. Higher amounts of rain will be added over the Olympic and Cascade mountains.
ALERT: A High Wind Watch for all of western Washington, Saturday afternoon and night, as a powerful storm is developing and threatens to bring dangerous wind speeds, including downtown cities.


Push back against darkness and despair . . .
"Push Back Against Despair! It's Time to Receive Your Portion"
The Father says today, "As you press into the pressure you will break out into the Kingdom. My Kingdom," says the Father, "is motion activated." This is your time to take action. It doesn't matter what the opposition is doing or saying because their plans do not determine what happens next in your life. I am this very day bringing you out of the captivity—brought on by putting your attention on what the enemy is thinking and planning.
"My plans for you are life and life more abundantly. My plans are to give you a hope and an expected end. My plans are to pour out upon you, even this day, all things that pertain unto life and godliness as you fill your heart and mind with the knowledge of what I am doing, regardless of the attempt of the enemy to distract you."


The Community of Pelzer Historical Society (CPHS) has graciously allowed us to reprint an article that appeared on their website in January of 2010. Please take a few minutes to visit their website at www.historicpelzer.org and if you'd like more information about that area of South Carolina, contact Beth at beth@historicpelzer.org.
Late in the afternoon of August 15, 1591, two small ships sailed into the bay at Hatorask (Hatteras)and dropped anchor some three leagues from the shore. As twilight fell one of the voyagers paced the deck restlessly from stern to stern. The voyager was John White, Governor of Sir Walter Raleigh's Roanoke colony, who was finally returning to the little band of English settlers, among them his own daughter, whom he had left nearly four years before to return to England for supplies. Upon first coming to anchor in the place where the Colony were left in 1587, John and the others saw .."..a great smoke..". This had them in high hopes that all was well and that the colonists were in John's words.."…there expecting my return out of England." John found no sign of the colonists at the site. An agreement had been established between John's group and the colonists in which in the event that the colonists were to run into any trouble, they would .."carve over the letters or name a Cross in this form (a Maltese cross with arms of equal length..) There were no such signs of distress. After exploring at least two signs of smoke, which led to no positive result, John White and his group closely observed at the site "…in all this way we saw in the sand the print of the Savages' feet of 2 or 3 sorts…and as we entered up the sandy bank, upon a tree, in the very brow thereof, were curiously carved these fair Roman letters CRO: which letters presently we knew to signify the place where I should find the planters seated, according to a secret token agreed upon between them and me at my last departure from them; which was, that in any ways they should not fail to write or carve on the trees or posts of the doors the name of the place where they should be seated; for at my coming away they were prepared to remove from Roanoak 50 miles into the main." White was disturbed to find that the houses had been taken down and in their place built a "..high palisade much like a fort…". After closer inspection, White found "..one of the chief trees or posts at the right side of the entrance had the bark taken off, and five foot from the ground in fair Capital letters was graven CROATOAN without any cross or sign of distress.." There were a few items found and the area was nearly overgrown with grass and weeds. John White and his men ended up sailing back to England and any other search was abandoned. Thus, the Roanoke Colony became the famous "Lost Colony" in American history. It should be remembered that this colony included the first child born of English parents in the new world, Virginia Dare, daughter of Eleanor, herself the daughter of John White, and Ananias Dare.
One day, in 1937, a tourist found a stone with what appeared to be Elizabethan English words on the banks of the Chowan River near Edenton, North Carolina. Eventually this stone found its way to the desk of a professor at Emory University, by the name of Haywood J. Pearce Jr., an authority on fourteenth to seventeenth century English writing. He aroused the interest of his father, Dr. H J Pearce Sr. , then President of Brenau College in Gainesville, GA. Then, in 1939, a "Bill" Eberhart, of Fulton County, GA was traveling by automobile through the upcountry of South Carolina. He pulled over to take a nap in his car and realized that he had a flat tire. He used a stone that he picked up in a clay ravine to help raise his car from the ground. He noticed the stone had writing on it. He was curious and went back to the ravine and found twelve more similar stones. The site of Eberhart's discovery was a hillside on the Greenville County side of the Saluda River some twelve miles below Greenville just outside the town of Pelzer on Hwy 20. The story told from these stones is indeed amazing as it tells of a 350 mile trek begun by one hundred and seventeen settlers of the Roanoke Colony to the southwest through North Carolina into South Carolina. By the time they arrived near the hillside by the Saluda River, their number had diminished to twenty four. At this place, the band was attacked by savages, resulting in seventeen additional persons killed, including Ananias and Virginia Dare. The seventeen were then buried on that hillside.
It is said that WWII precluded further study of the stones and that Dr. Pearce Sr. died in 1943. Additionally, his heirs sold the Pelzer hill to P. M. McClane for $700. Although no conclusive statement can be made about the stones, is it possible that the blue-eyed Lumbee Indians of the Laurinburg area of North Carolina, and the bearded Keyauwee Indians who lived near the Chattahoochee River, could have acquired these characteristics from intermarriage with remnants of the Roanoke Colony?
This entry was posted in Croatoan, Keyauwee, Lost Colony, Lumbee. Bookmark the permalink.
Very close to home Roberta, thanks ! This area is full of artifacts that have yet to be discovered. My mother told me about living in North Greenville beside a pasture that was once an old Indian camp. She and her siblings would play there as children and found several Native tools and arrowheads. I asked her why she didn't take any of them home and save them. She said, "they were just old knick-knacks, what were we going to do with them ?" I think I just looked at her for a moment in complete silence after that !


The fire hydrant meter program provides service for customers within the City of Durham and outlines specific procedures to follow for authorized use of public fire hydrants. According to the Durham City Code, only two organizations, the City of Durham Fire Department and select Divisions of the Department of Water Management are authorized to use fire hydrants without a special permit. Private contractors and individuals must apply for access to public fire hydrants.
Every day businesses such as street cleaners, pressure washers, and those working on construction projects need to use water from a fire hydrant. The City of Durham Water and Sewer Maintenance Division offers these customers a permit for a Hydrant Meter Assembly to obtain water for non-drinking use in a safe and legal manner.
The Water and Sewer Maintenance Division will provide temporary water service by means of a public fire hydrant to customers who qualify for this service at locations within the service area. The provided Standard Fire Hydrant Meter Assembly offers a 2 ½-inch NST fire hose connection and a 3/4 MIPT garden hose connection, while the Jumper Box Assembly offers a 2" inlet and outlet connection for soft copper. Jumper Boxes are typically used when a direct tap is needed on a jobsite and no local fire hydrants are available. Individuals or companies must apply for access to a hydrant for non-drinking water use.
Applicants must provide the desired fire hydrant location which shall include unobstructed access to the job site. The fire hydrant's location and condition may affect availability and authorization of service. If the desired fire hydrant is deemed unsatisfactory for a temporary water connection Water and Sewer Maintenance staff will suggest the next best fire hydrant for providing temporary water service.
Additionally, The City of Durham Water and Sewer Maintenance Division reserve the right to deny service to any customer depending on equipment availability, water use restrictions, weather conditions or system capabilities. The City of Durham Water and Sewer Maintenance Division will grant access to a Fire Hydrant Meter Assembly for temporary water service only to customers who have satisfied the requirements for obtaining such services.
Call 919-560-4344 and select option #4 to inquire about a temporary Fire Hydrant Connection Service and availability.
Obtain the hydrant meter assembly permit application (PDF).
Fees must be paid in person at the cashiers counter located 101 City Hall Plaza.
Upon acceptance of the Hydrant Meter Application and necessary fees, Customer Billing Services Staff will create and submit an internal Service Request to install a Hydrant Meter assembly.
Upon processing of the Application and Service Request the customer will be contacted by Water and Sewer Maintenance staff to schedule installation of the Fire Hydrant Meter Assembly.
In most cases the Fire Hydrant Meter Assembly can be installed within five business days of payment.
City of Durham WSM Staff will contact Customer to schedule the installation of the Fire Hydrant Meter Assembly. Installation is available between 7:30am and 2:30 pm.
The City of Durham Water and Sewer Maintenance Division (WSM) will install and secure a Fire Hydrant Meter Assembly on the requested fire Hydrant if possible. If the requested hydrant is unfit to accept a Fire Hydrant Meter Assembly WSM staff will suggest and install the equipment at the next best location.
WSM staff will install and secure the Fire Hydrant Meter Assembly to and turn on the fire hydrant and ensure all equipment is properly working before leaving the site.
The customer will have access to and only be allowed to operate the 2" hand valve located on the exterior of the enclosed Fire Hydrant Meter assembly. Using a personal lock is permissible to secure the 2" hand valve.
All Fire Hydrant Meter Assemblies include freeze protection to ensure proper and continuous operation throughout the year.
The Jumper Box Assembly offers a 2" inlet and outlet connection for soft copper to be used when a direct tap access to water is needed.
The customer is responsible for immediately reporting any damage or concerns to the Fire Hydrant Meter Assembly to the Division of Water and Sewer Maintenance.
When the Customer is finished with the Fire Hydrant Meter assembly they must contact the Customer Billing Services division at 919-560-1200 at least two business days in advance to schedule retrieval of the equipment. Retrieval of equipment can be scheduled between 7:30 a.m and 2:30 p.m.
The City of Durham offers only secured Fire Hydrant Meter Assemblies for temporary water service. Vehicle mounted or portable assemblies are not an option.
If a customer wishes to have their Fire Hydrant Meter Assembly relocated, an advance notice of two business days shall be provided to Water and Sewer Maintenance staff and is subject to prior commitments. Hydrant Meter customers are not authorized to relocate the Fire Hydrant Meter Assembly.
The City of Durham shall complete an inspection and obtain a water consumption reading on a regular basis for Customers that are in possession of Hydrant Meter Assemblies for extended periods of time.
If the City of Durham discovers or determines that a customer is in violation of any portion of the Hydrant Meter Guidelines or associated policies, WSM Staff will immediately remove the Equipment and recommend the Customer's account and deposit be held until an investigation has been completed to determine if additional fees or penalties are required.
What are your sizes of hydrant meters?
What are your fees for hydrant meters?
How much water will I get from my meter?


As you read or hear Bible stories did you ever wish you had access to the 'backstory' – the interesting stuff that wasn't recorded in the official account? For example, when Simon (Peter) and his brother Andrew abandoned their fishing business to follow Jesus and become "fishers of men" (Mark 1:16-18) do you ever wonder what their wives had to say to them when they returned home? ("You've decided to fish for WHAT?") Wouldn't you really like to know how Jesus' host at the meal in Bethany acquired the name "Simon the Leper"? (Mark 14:3) Was this perhaps a lucky guy that Jesus had previously healed? Or was this just a real bummer of a nickname? And just what exactly was the deal with the young man dressed only in a cloak who slipped out of it in the garden of Gethsemane and ran away naked? (Mark 14:51-52) Who was he and why was he running around there in such an incomplete state of attire? (It had to have been an interesting story how he made it home in that condition.) Perhaps most perplexing of all: why was this oddball anecdote considered important enough for Mark to include in his otherwise somber passion account? Inquiring minds would love to know!
Yet we still wonder about all those people we read about. What were the reactions of the bride and groom at the wedding of Cana when they realized that their inadequate supply of wine had been miraculously supplemented? (John 2:1-11) One would like to think this became a "signature moment" of a long and happy marriage – an event they lovingly recounted to their children and grandchildren. Or were they perhaps too caught up in the excitement of the festivities to really appreciate what happened? (Or did the groom prudently keep mum with his new bride about his poor planning?) Or here's one: what was it like to be one of Jesus' kid brothers or sisters? (Mark 6:3). Were you always getting caught in the short end of a comparison: "Why can't you behave like your brother?" Really, wouldn't you like to be a mouse in the corner of that home?
Some might find it a bit uncomfortable to entertain such 'irreverent' speculations. Perhaps they grew up thinking that Bible stories transpired in a different kind of reality than we live in today. And surely that was the case: in terms of the specific physical circumstances they lived in, things were different in pretty much every possible way. Yet – these were still real people that had problems, feelings, and behaviors just like ours. They got hungry and tired, were desperately concerned about their sick children, grieved for loved ones who died. And we also empathize with their failings: they could be insensitive and argumentative, miss the point of a sermon, fall asleep at really inopportune times, and even abandon and deny the one that they knew to be the Son of God. Yup – we recognize them all too well!
The Bible is not a fairy tale about some kind of "once upon a time" fantasy world painted in pastel colors and populated like a Hollywood movie set by unrealistically pious and attractive people. No, it is a gritty account of real folks – people like us – struggling to find ultimate meaning in life while dealing with the challenges of everyday existence and relationships. It's the story of how God invaded our crazy world of fatally flawed and self-absorbed people and showed us the only way out. A way that was deceptively simple (just believe) but came with a terrible price – the torture and death of God's own beloved Son. That's a story of supernatural love that transcends anything in human experience. Yet it's anchored in earthly reality, as witnessed by the real human stories that fill the Bible – both those written and the untold ones that hover in the background.


Do you ever get nervous just thinking about going to the dentist? If you're tense or anxious, let us know before your appointment. Getting your concerns out in the open will help us adapt the treatment to your needs.
Try to choose a time for your dental visit when you're less likely to be rushed or under pressure. If the sound of the drill bothers you, bring a portable audio player and headset so you can listen to your favorite music. If you'd like to take a mild sedative before your appointment or if you prefer to be asleep during your treatment please let us know.
What should every patient know about dental anesthesia?
Advances in medications, dental anesthetics and anesthesia techniques can greatly reduce even eliminate discomfort during dental treatment. The following explains options available to help alleviate anxiety or pain that may be associated with dental care.
Topical anesthetics are applied to mouth tissues with a swab to prevent pain on the surface level in preparation for administering an injectable local anesthetic.
Injectable local anesthetics, such as Lidocaine, prevent pain in a specific area of your mouth during treatment by blocking the nerves that sense or transmit pain and numbing mouth tissues. They cause the temporary numbness often referred to as a "fat lip" feeling. There are several types of local anesthetics, allowing us to select the one most suited to your needs. Local anesthetics are always used in endodontic procedures.
Anti-anxiety agents may help you relax during dental visits and are used along with local anesthetics. Dentists also can use these agents to induce "conscious sedation," in which the patient achieves a relaxed state during treatment while remaining responsive. Sedatives are usually taken by mouth before dental procedures.
When patients have high levels of anxiety or when they simply want the maximum level of comfort during their treatment, we able to provide "sleep dentistry." Intravenous anesthetics produce a deeper level of sedation that eliminates all anxiety and sensation. There the patient retains normal body functions, and the recovery time is relatively short.
This method of deep sedation or general anesthesia is administered in our office by a board certified anesthesiologist, and may be recommended in certain procedures for patients with severe anxiety or a history of resistance to local anesthetics, small children, or for those who prefer ultimate comfort.
Dentists use the pain and anxiety control techniques mentioned above to treat tens of millions of patients safely every year. Even so, taking any medication involves a certain amount of risk. That's why we urge you to take an active role in your oral health care. This includes knowing your health status and telling us about any illnesses or health conditions, whether you are taking any medications (prescription or non-prescription), and whether you've ever had any problems such as allergic reactions to any medications. It also includes understanding the risks and benefits involved in dental treatment, so that together with you we can make the best decisions about the treatment that is right for you.
Understanding the range of choices that are available to relieve anxiety and discomfort makes you a well-informed dental consumer. If you have questions or concerns about your oral health care, don't hesitate to call us. If you still have concerns, consider getting a second opinion. Working together with you, we can choose the appropriate steps to make your endodontic treatment as safe and comfortable as possible.


Fans have learned that Fox rejected an episode of Batman: The Animated Series that focused on Black Canary and Catwoman because the network wanted more Robin. Widely regarded as one of the best superhero animated shows of all time, Batman: The Animated Series ran for 85 episodes on Fox Kids between 1992 and 1995. Bruce Timm and Eric Radomski developed the series.
Today's news comes from Paul Dini, who served as a writer on Batman: The Animated Series and is currently trying to encourage his fans to invest in his Kickstarter-funded Boo & Hiss graphic novel. Dini's most famous creation, Harley Quinn, has also been in the news recently: it was recently announced that DC is developing a Harley Quinn animated show for Warner Bros' upcoming online streaming service. Somewhat weirdly, it doesn't look like Paul Dini and Bruce Timm – who created Harley together for Batman: The Animated Series – are involved with that project at all.
It sounds like Fox weren't willing to let Dini ditch Robin for a week so he could focus an episode on Black Canary (who ultimately never appeared in the series) and Catwoman (who showed up several times and was voiced by Adrienne Barbeau). It's a shame, particularly because a female double-hander would have been quite forward thinking for the time.
However, this isn't the only idea that Dini didn't manage to see to fruition. There was also "a Poison Ivy seducing Swamp Thing story" that couldn't work because "the rights to Swampy weren't available then." As Dini notes, he recycled that idea and "did a sort of version of it years later as an episode of Justice League Action." Maybe, one day, the Catwoman/Black Cat idea will find a place on another DC series, too.
Keep it Screen Rant for all the latest DC news.


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Ethiopia mediates Gedo leadership conflict in Somalia
Staff reporter GAROWE ONLINE
Posted On 23-11-2022, 04:38PM
MOGADISHU, Somalia - Authorities in Ethiopia successfully mediated a leadership dispute in the Gedo region of Somalia, it has now emerged, months after federal authorities in the country clashed with local leaders in Jubaland, and was almost degenerated into bloodshed.
The government of Somalia then under the stewardship of Mohamed Abdullahi Farmajo deployed elite soldiers to the Gedo region where local leaders loyal to regional leader Ahmed Islam Mohamed Madobe were dethroned, leading to endless conflicts which almost dismantled coordination.
Reports from Gedo indicate that a number of officials from the Somali region of Ethiopia crossed over the border where they mediated differences between regional Governor Ahmed Bulle Gared and spy chief Bakal Koke, who have been tussling over sharing revenue from road taxations in the region.
Taxation in Somalia is not centralized given years of civil war thus lack of coordination between the national and regional governments. In fact, even the Al-Shabaab militants manage to collect colossal amounts of money from the locals, which helps them to finance their illegal activities.
In a ceremony held in Dolow on Wednesday, both officials promised to work together and end internal strife hampering the administration's ability to deliver essential services to residents in the region. Gedo region is strategic to Ethiopia's interests in Somalia.
For a long time, the Ethiopian authorities have been using Gedo as a buffer zone in the fight against Al-Shabaab, with the most recent being in August when Al-Shabaab raided Eastern Ethiopia. Authorities in Addis Ababa reported that Somali regional troops and Ethiopian forces managed to neutralize the group.
To improve the security situation in the country, Ethiopia has since managed to establish the buffer zone and Wednesday's decision to reconcile local leaders is believed to be a strategy to further neutralize infiltration of Al-Shabaab, who usually take advantage of leadership disputes to commit crimes.
Although details about the meeting remain scarce, the Ethiopians just like their counterparts from Kenya have for a long time been fighting for the stability of Jubaland which borders them. Both Kenyan and Ethiopian troops are stationed in various towns across Jubaland where they have managed to dislodge Al-Shabaab.
GAROWE ONLINE