The liver is an important multifunctional organ that has a remarkable capacity to regenerate. Any acute or chronic liver injury can result in liver regeneration. Following liver injury, most hepatocytes exit G0 phase and re-enter the cell cycle to generate new hepatocytes. Liver regeneration is an orchestrated process that is regulated by multiple inflammatory cytokines, growth factors and hormones. Inflammatory cytokines, such as IL-6 and TNFα, play critical roles in the initiation of liver regeneration. During liver regeneration, IL-6 binds to its receptor, IL-6R, and further activates signal transducer and activator of transcription (Stat) 3 signaling pathways, resulting in increased hepatocyte survival and proliferation [1, 2]. By activating NF-κB, TNFα also facilitates the priming of hepatocyte proliferation [3, 4]. However, the mechanism by which the production of these inflammatory cytokines is regulated during liver regeneration remains elusive.
Despite of the potent regenerative capacity of the liver, nearly one million deaths per year are associated with chronic liver failure, indicating that, in these cases, liver regeneration was not sufficient to compensate for the loss of hepatocytes or to restore liver function. Therefore, novel therapeutic targets for enhancing liver regeneration are definitely desired by those patients suffering from fatal liver injuries.
Osteopontin (OPN), which is also known as secreted phosphoprotein 1 (Spp1) and early T-lymphocyte activation 1 (Eta-1), is a multifunctional protein that is involved in hepatic inflammation and diverse liver diseases [5, 6]. OPN is significantly induced in macrophages after stimulation by cytokines such as IL-6, TNFα, IL-1β and IFNγ [7, 8]. Interestingly, OPN can function as a ligand to induce the secretion of IL-12 and TNFα from peritoneal macrophages . In addition, OPN has been suggested as a mediator for macrophage activation, migration and infiltration [9, 10]. It has been reported that OPN promotes the migration of Kupffer cells isolated from normal rats, suggesting that OPN may act as a chemoattractant to recruit macrophages in the liver . Earlier studies indicated that the absence of OPN decreased macrophage infiltration in the liver after liver injury [12, 13]. However, whether OPN modulates liver regeneration by regulating liver inflammation or macrophage infiltration has not been determined.
The aim of this study was to investigate the role of OPN in liver regeneration and to understand the underlying mechanisms. In the present study, we demonstrate that OPN is an important regulator that facilitates the priming phase of liver regeneration following PHx. The advantages conferred OPN during liver regeneration are attributed to its functions in enhancing hepatic recruitment of macrophages and neutrophils, inducing IL-6 production, and subsequently promoting Stat3 activation in hepatocytes.
All animal studies were approved by the Institutional Animal Care and Use Committees of Renji Hospital Clinical Stem Cell Research Center and Shanghai Jiao Tong University. OPN knockout (OPN-/-) mice (B6.Cg-Spp1tm1Blh/J) and their wild-type (WT) control C57BL/6J mice were purchased from the Jackson Laboratory (Bar Harbor, ME).
Eight- to ten-week-old male OPN-/- and age-matched WT mice were subjected to two-thirds partial hepatectomy (PHx) as previously described . Briefly, after anesthetization, the left lateral lobe and the median lobe plus gall bladder were removed. The mice were sacrificed at various time points after surgery. Mice were injected intraperitoneally with BrdU (Sigma-Aldrich, St. Louis, MO) at 50 mg/kg body weight 2 hours before sacrificed. The regenerated liver was weighed, and the liver-body weight ratio was calculated. After anesthetization, both liver tissues and blood plasma from the portal vein 3 hours after sham surgery or PHx were collected and analyzed for endotoxin using an endotoxin test kit (Houshiji, Xiamen, China) according to the manufacturer's protocol. Serum was collected for tests, including alanine transaminase (ALT) and aspartate aminotransferase (AST) analyses using microplate test kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer's protocols. The regenerated liver tissues were collected for molecular and biochemical analyses.
The livers were harvested and fixed in 4% paraformaldehyde for at least 24 hours. Liver samples, embedded in paraffin, were cut into 5-μm-thick sections. Liver tissue sections were stained with hematoxylin-eosin (H&E) using standard procedures. For immunohistochemistry, sections were rehydrated and processed for an antigen-unmasking procedure, then incubated with primary antibodies overnight at 4 °C, followed by horseradish peroxidase-conjudated secondary antibodies. For all tissues, sections were counterstained with hematoxylin. Six to ten images from random fields in each section were taken with a light microscope (Axio Imager A1; Zeiss), and at least three mice per group were subjected to each experiment. Image-Pro Plus 6.0 was used for image analysis of sections. The primary antibodies used were goat anti-OPN (R&D Systems, Minneapolis, MN), mouse anti-BrdU, mouse anti-PCNA (Cell Signaling, Boston, MA), rabbit anti-MPO (BioCare Medical, Concord, CA), and rat anti-F4/80 (AbD Serotec, Oxford, UK).
Liver tissues and cell samples were homogenized in RIPA buffer (Thermo Scientific, Rockford, IL) containing a protease inhibitor cocktail (Calbiochem, Raleigh, NC). Protein concentrations were quantified with a BCA protein assay kit (Thermo Scientific, Rockford, IL) according to the manufacturer's manual. Fifty micrograms of total protein from each tissue or cell lysate sample were loaded and separated by gel electrophoresis and then transferred to nitrocellulose membranes. After blocking, membranes were incubated with primary antibodies at 4 °C overnight under shaking conditions. The membranes were then incubated with horseradish peroxidase-conjugated secondary antibodies. Detection and quantification of protein bands were performed using the ChemiDocTM XRS+ System with Image LabTM Software (Bio-Rad). The primary antibodies used were rabbit anti-phospho-Stat3 (Tyr705), rabbit anti-Stat3, rabbit anti-phospho-IκBα (Ser32), mouse anti-IκBα (Cell Signaling), anti-β-actin (Sigma).
Total RNA was purified from approximately 30 mg liver samples or cell lysates according to the manufacturer's protocol (Qiagen, Valencia, CA), and then 500 ng mRNA was reverse-transcribed to cDNA using a RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific, Rockford, IL). Relative quantitative gene expression was measured via real-time PCR using a ViiATM 7 Real-Time PCR System (Life Technologies) and a SYBR Green PCR Kit (TaKaRa Biotechnology, Dalian, China). GAPDH expression was used as an internal standard. The primer sequences are as follows (5'-3'): GAPDH sense: AGGCCGGTGCTGAGTATGTC, anti-sense: TGCCTGCTTCACCACCTTCT; PCNA sense: GTGGAGCAACTTGGAATCCC, anti-sense: GGTTACCGCCTCCTCTTCTT; IL-6 sense: TGTTCTCTGGGAAATCGTGGA, anti-sense: TTTCTGCAAGTGCATCATCGT; TNFα sense: TTCTATGGCCCAGACCCTCA, anti-sense: TTTGCTACGACGTGGGCTAC; MCP-1 sense: CCCTAAGGTCTTCAGCACCT, anti-sense: ACTGTCACACTGGTCACTCC; F4/80 sense: TCTGCAGTGTCAGCTCAGAA, anti-sense: GTATGCCATGATGCTTGCCA; MPO sense: CTCGATGGAATGGGGAGAAGC, anti-sense: CCAACACCAAGGGCAGGTAGTC; c-fos sense: TACTACCATTCCCCAGCCGA, anti-sense: GCTGTCACCGTGGGGATAAA; c-jun sense: CGATGCCCTCAACGCC, anti-sense: CTTAGGGTTACTGTAGCCGTAGGC; c-myc sense: ATGTTGCGGTCGCTACGTC, anti-sense: AGAAGTTGCCACCGCCG; HGF sense: CAAGCAATCCAGAGGTACGC, anti-sense: CCATGGGACCTCTGTAGCTT; IL-22 sense: CAGGTTCCAGCCCTACAT, anti-sense: CAGCCTTCTGACATTCTTCT; and IL-22Rα1 sense: TTCCTCGGCTCCATCACA, anti-sense: AGCGTCTTCACTCGGCAC.
Liver leukocytes were isolated as previously described  from livers of mice with minor modifications. In brief, liver tissues were passed through a 70-μm cell strainer (BD Falcon) in Hanks' balanced salt solution. The cell suspension was centrifuged at 500 ×g for 10 minutes. The cell pellet, including hepatocyte debris, was re-suspended in 5 ml of 35% Percoll (GE Healthcare Life Science), and the cell suspension was centrifuged at 500 ×g for 15 minutes at room temperature. The cell pellet containing the leukocytes was harvested and re-suspended in 5 ml of ACK Lysing Buffer (Gibco) to remove red blood cells. After incubated for 5 minutes, cells were washed twice and counted. The resulting leukocytes were incubated with APC-conjugated anti-CD45, PE-conjugated anti-CD11b, PerCP Cyanine5.5-conjugated anti-F4/80 (eBioscience, San Diego, CA), FITC-conjugated anti-Gr-1 (Miltenyi Biotec, Bergisch Gladbach, Germany) and analyzed using a BD FACSCalibur flow cytometer. The purity of the isolated leukocytes was over 90%. The number of neutrophils and macrophages per liver was calculated by multiplying the percentage of each population from the flow cytometric data by the total number of leukocytes per liver.
Primary cells were isolated by perfusion of the liver with Gey's balanced salt solution containing collagenase (Sigma). In brief, the liver was perfused via the portal vein at 5 ml per minute with EGTA buffer for 8 minutes. Subsequently, the liver was perfused with perfusion buffer containing collagenase at 5 ml per minute for 10 minutes. The liver was then dissociated in digestion buffer containing collagenase and filtered with a 70-μm cell strainer. Hepatocytes were collected by centrifugation at 400 rpm for 5 minutes. The supernatant containing the non-parenchymal cells was further cleared by another centrifugation at 400 rpm for 5 minutes. Non-parenchymal cells were pelleted by centrifugation at 650 ×g for 10 minutes. HSC were purified by a 12.5% OptiPrep gradient (Sigma) and collected for analysis. Kupffer cells were purified by gradient centrifugation using 17% OptiPrep. The cell fraction including Kupffer cells was gently aspirated and plated in non-collagen coated culture plates for 20 minutes. Kupffer cells attached to the plate were collected for analysis. The purity of isolated Kupffer cells or HSCs was over 90%.
The isolated hepatocytes were counted and seeded in culture plates. After cell spreading, the culture medium was deprived of fetal bovine serum for 24 hours and cells were subjected to proliferation analysis in the presence of 50 nM of recombinant mouse OPN (rmOPN) (R&D Systems) stimulation according to dose-response (5-500 nM) experiments. Cells were then subjected to immunofluorescence for PCNA after 48 hours of treatment, and cell lysates were collected for PCNA mRNA analysis following 24 hours of treatment. Isolated Kupffer cells were cultured in RPMI 1640 medium supplemented with 10% FBS (Gibco). After spreading overnight, cells were starved and treated with 100 nM of rmOPN for the indicated time according to dose-response (5-500 nM) experiments. Cell lysates were collected for mRNA analysis and cell supernatants were collected for ELISA analysis.
Frozen liver sections (5-μm thick) and cells were fixed. Nonspecific antibody binding was blocked by 5% goat serum and 0.5% Triton X-100 in PBS. After aspirating away the blocking buffer, cells were incubated with mouse anti-PCNA, rabbit anti-desmin (Cell Signaling, Boston, MA), goat anti-OPN (R&D Systems), rabbit anti-CK19 (Proteintech, Chicago, IL), or rat anti-F4/80 (AbD Serotec) antibody diluted in PBS containing 1% bovine serum albumin and 0.5% Triton X-100 overnight at 4 °C. After washing in PBS, cells were incubated with fluorescence-conjugated secondary antibodies at room temperature for 1 hour. DAPI was used to stain cell nuclei. Six to ten images were acquired for each sample and the percentage of positive nuclei was calculated.
ELISA kits were used to determine liver and serum OPN levels (USCN Life Science, Wuhan, China) and IL-6 and TNFα levels in the serum and the cell supernatants (NeoBioscience Technology, Shenzhen, China) according to the manufacturer's protocols.
Results are expressed as the mean ± SEM. The results were assessed using two-tailed, unpaired or paired Student's t-tests. P<0.05 was considered statistically significant (*=P<0.05/**=P<0.01/***=P<0.001).
To investigate the role of OPN in liver regeneration, we first measured OPN protein levels in the serum and liver after PHx. OPN levels significantly increased in both the serum and liver after 3 hours and reached the peak at 12 hours post-PHx (Fig. 1A and B). We then sought to determine which cell type contributed to OPN production during liver regeneration. In WT adult mouse livers, consistent with previous studies [12, 16], OPN was selectively expressed in epithelial cells around bile ducts (Fig. 1C). A significant increased staining intensity was observed for OPN in these epithelial cells early after PHx (Fig. 1C). To further define these cells, double immunofluorescence staining with anti-OPN and anti-F4/80 (a specific marker for macrophages), anti-desmin (a specific marker for HSCs) or anti-CK19 (a specific marker for cholangiocytes) antibodies was performed. There was no co-localization between OPN and markers of Kupffer cells or HSCs (Fig. S1A and B), suggesting that OPN is not expressed by these cell types after PHx. Interestingly, CK19 and OPN were highly co-localized 3 hours after PHx (Fig. 1D). Although CK19 also serves as a marker of oval cells, given that oval cells were usually absent from the normal liver controls and are normally activated 5-10 days after PHx [17, 18], we concluded that the CK19-positive cells that were labeled at 3 hours after PHx were mainly BECs (Fig. 1D).
OPN knockout (OPN-/-) mice were used to determine the in vivo function of OPN during liver regeneration. There was no difference in liver weight, body weight or the liver-body weight ratio between 8-week-old WT and OPN-/- mice, suggesting that depletion of OPN did not impact liver growth in adult mice (Fig. S2A). Histological analyses also revealed that the liver from OPN-/- mice displayed normal histology (Fig. S2B).
We then performed two-thirds partial hepatectomy in both WT and OPN-/- mice and collected sera and liver tissues at the indicated time points after PHx. Serum ALT and AST levels were greatly increased at 12 hours post-PHx in both WT and OPN-/- mice, but no significant difference in serum ALT or AST levels was observed between WT and OPN-/- mice (Fig. 2A and B). BrdU incorporation assays indicated that WT mice displayed peak hepatocyte proliferation at 40 hours after PHx, but the proliferative peak in OPN-/- mice was delayed to 48 hours after PHx (Fig. 2C). Similar results were obtained by staining with PCNA, which is a marker for proliferating cells (Fig. 2D). In addition, the difference in the liver-to-body weight ratio between WT and OPN-/- mice was significant at 40 hours after PHx, but the ratio became comparable at 48 hours after PHx (Fig. 2E). These findings indicate that compared to WT, OPN-/- mice exhibit delayed liver regeneration after PHx.
To uncover the mechanism by which OPN depletion causes delayed hepatic proliferation, we examined whether OPN has a direct effect on hepatocyte proliferation. Freshly isolated hepatocytes were treated with recombinant mouse OPN (rmOPN) and the proliferative activity of hepatocytes was determined by measuring PCNA levels. As shown in Fig. 3A, comparable PCNA mRNA levels were detected in hepatocytes that were treated with rmOPN at the indicated concentrations. In addition, there was no difference in PCNA-positive hepatocytes between the control and rmOPN-treated groups (Fig. 3B and C). These results suggest that OPN does not induce hepatocyte proliferation directly.
OPN levels increase in the serum and regenerating liver after PHx. (A) Serum and (B) liver OPN levels were determined by ELISA at the indicated time points after PHx in WT mice (n = 4-10). The data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 compared with 0 hour. (C) Immunohistochemistry of OPN in livers from WT mice and livers at 3 and 12 hours after PHx (200× magnification). At least three experiments were performed with similar results. (D) Immunofluorescence analysis showed co-staining of OPN and CK19 at 3 hours after PHx (200× magnification). Three independent experiments were performed with similar results.
OPN deletion delays the peak of hepatocyte proliferation during liver regeneration after PHx. (A) Serum ALT and (B) AST activities were determined at different time points after PHx (n = 3-8). The data are shown as the mean ± SEM. (C) BrdU incorporation were measured in liver sections at the indicated time points after PHx (200× magnification). Quantitative analysis of BrdU labeling was performed (n = 3-7). The data are shown as the mean ± SEM. *P<0.05. (D) PCNA immunolabeling in liver sections after PHx (200× magnification). The number of PCNA positive cells in remnant liver lobes was counted in at least 5 fields per liver section (n = 3-7). The data are shown as the mean ± SEM. *P<0.05, **P<0.01. (E) The liver-to-body weight ratio was determined at the indicated time points after PHx (n = 3-8). The data are shown as the mean ± SEM. *P<0.05.
OPN does not stimulate hepatocyte proliferation in vitro. Primary hepatocytes isolated from WT mice were treated with rmOPN. (A) Hepatocyte lysates were collected 24 hours after OPN treatment and analyzed for PCNA mRNA expression (n = 3). The data are shown as the mean ± SEM. (B) Immunofluorescence for PCNA was performed 48 hours after treatment with 50 nM rmOPN (200× magnification). Three independent experiments were performed with similar results. (C) Quantitative analysis of PCNA immunolabeling was performed (n = 3). The data are shown as the mean ± SEM. RFC: relative fold change.
Previous studies have reported that OPN is a key regulator of inflammatory processes, which indicates that OPN is not only considered to be a pro-inflammatory cytokine but also a potent chemoattractant for macrophages and neutrophils [19, 20]. Thus, to further investigate the association between OPN deletion and delayed liver regeneration, we studied the correlation between OPN and the inflammatory response in regenerating livers. FACS analysis of liver immune cells was performed at 3 hours post-PHx in WT and OPN-/- mice. Compared to WT mice, OPN-/- mice showed significantly impaired hepatic recruitment of macrophages and neutrophils 3 hours after PHx (Fig. 4A, Fig. S3A and B). Reduced hepatic recruitment of macrophages and neutrophils in OPN-/- mice was also confirmed by immunohistochemical staining and qPCR at the indicated time points post PHx (Fig. 4B-E). Consistent with a reduction in hepatic recruitment of macrophages and neutrophils, impaired induction of MCP-1, also known as monocyte chemotactic and activating factor, was also observed in the livers of OPN-/- mice 3 hours after PHx (Fig. S3C). Interestingly, compared to WT mice, although the hepatic recruitment of macrophages was restored, the liver infiltration of neutrophils was maintained at a very low level in OPN-/- mice 12 hours after PHx (Fig. 4B and C), suggesting that the hepatic recruitment of these two cell types was modulated by different chemokines. Given the critical roles of IL-6 and TNFα in the initiation of liver regeneration after PHx, we then measured the protein and mRNA levels of those two cytokines in the serum and liver. Compared to WT mice, OPN-depleted mice showed a significant reduction in IL-6 at 3 hours after PHx (Fig 5A and B). Although significantly reduced induction of TNFα mRNA was observed in the liver in OPN-/- mice compared to WT mice at 3 hours after PHx, we failed to confirm this difference at the protein level in serum (Fig. S4A and B).
Kupffer cells are the major source of IL-6 during liver regeneration . Although impaired hepatic recruitment of macrophages could contribute to reduced IL-6 levels in OPN-/- mice after PHx, other possibilities should not be ruled out. One possibility is that OPN might directly drive Kupffer cells to produce IL-6, and the other is that Kupffer cells in WT mice are more activated than those in OPN-/- mice after PHx. We isolated Kupffer cells from WT livers and treated them with 100 nM rmOPN. Quantitative PCR and ELISA assays were performed to examine changes in IL-6 at the mRNA and protein levels respectively. rmOPN treatment slightly increased IL-6 production but significantly stimulated TNFα expression in Kupffer cells (Fig. 5C and D, S4C and D), suggesting that the direct function of OPN on Kupffer cells may not attribute to reduced IL-6 levels in OPN-/- mice post-PHx. To examine the in vivo status of Kupffer cell activation, we isolated Kupffer cells, HSCs and hepatocytes from the livers of WT and OPN-/- mice after PHx at the indicated time points. IL-6 levels were measured by qPCR. Compared to WT counterparts, Kupffer cells from OPN-/- mice produced significantly less IL-6 at 3 hours after PHx (Fig. 6A). No induction of IL-6 was observed in either the HSCs or the hepatocytes after PHx (Fig. 6A). Similarly, Kupffer cells from WT mice produced a greater amount of TNFα than those from OPN-/- mice 3 hours after PHx (Fig. S4E). These results indicate that Kupffer cells are more activated in WT mice compared to OPN-/- mice 3 hours after PHx and suggest that the decrease in serum IL-6 levels in OPN-/- mice after PHx is due to both impaired liver infiltration of macrophages and the reduced activation of Kupffer cells.
Endotoxin has been reported as an essential factor that contributes to the initiation of liver regeneration by binding to TLR4 to activate intracellular signaling pathways, which finally induces Kupffer cells to produce IL-6 and TNFα [4, 22, 23]. To answer why WT Kupffer cells were more activated than OPN-/- cells after PHx, we measured the levels of gut-derived endotoxin in portal circulation and the liver. We found that the concentrations of hepatic and plasma LPS were significantly higher in WT mice compared to OPN-/- mice 3 hours after PHx (Fig. S5A and B), suggesting that different LPS concentrations may contribute to the differential activation of Kupffer cells in WT and OPN-/- mice after PHx. Because LPS can activate the NF-κB pathway in Kupffer cells, resulting in the production of IL-6 and increased phosphorylation of IκBα, indicating the activation of the NF-κB pathway , we then examined phosphorylated IκBα levels in both WT and OPN-/- Kupffer cells. As expected, phosphorylation of IκBα was significantly impaired in OPN-/- Kupffer cells compared to WT counterparts 3 hours post-PHx (Fig. S5C). These results clearly indicate that OPN indirectly affects Kupffer cell activation by modulating LPS release into portal circulation after PHx.
The Stat3 pathway plays a critical role in liver regeneration by promoting hepatocyte survival and liver repair. Given the key role of IL-6 in activating the Stat3 pathway, we then measured indicators of the Stat3 signaling pathway in hepatocytes after PHx. In response to reduction in IL-6 induction in OPN-/- mice, significant decreased Stat3 phosphorylation was observed in OPN-/- liver tissues compared to WT tissues (Fig. 6B). IL-22 has also been shown to play an important role in promoting liver regeneration by activating Stat3 . We quantified the mRNA expression of IL-22 and IL-22Rα1 by qPCR. Three hours after PHx, the IL-22 mRNA level was increased 1.5-fold in WT livers and no IL-22 induction was observed in OPN-/- livers (Fig. S6A). No induction of IL-22Rα1 was detected in either WT or OPN-/- mice 3 hours after PHx (Fig. S6B). Compared to OPN-/- mice, WT mice presented 1.5-fold and 10-fold higher IL-22 and IL-6 levels respectively 3 hours after PHx (Fig. 5B, Fig. S6A). Therefore, it seems that IL-6, but not IL-22, plays the primary role in activating Stat3 in our model.
To further confirm that OPN depletion impairs the activation of Stat3, the expression of the downstream genes c-fos, c-myc and c-jun was measured. As shown in Fig. 6C, all three of these immediate early genes were markedly higher in WT livers compared to OPN-/- livers 3 hours after hepatectomy. Given that HGF, a mitogenic and anti-apoptotic factor, is synthesized in response to IL-6 in the liver [26, 27], we then measured HGF levels in the liver 3 hours after PHx. We found that the level of HGF in WT mice was 1.5-fold higher than that in OPN-/- mice (Fig. S7). Collectively, these results indicate that by impairing hepatic recruitment of macrophages and blunting Kupffer cell activation, the loss of OPN consequently reduces IL-6 production and in turn attenuates the activation of the Stat3 pathway to impair the priming phase of liver regeneration.
Although a number of studies have suggested that OPN plays important roles in liver fibrosis, alcoholic hepatitis, non-alcoholic fatty liver disease and hepatocellular carcinoma [28-31], thus far, its role in liver regeneration has not been determined. The close relationship between OPN and hepatology implies that OPN, a multifunctional cytokine, may act as a regulator during liver regeneration. In this study, we identified OPN act as a novel positive regulator in liver regeneration. In the case of PHx, depletion of OPN significantly delayed the proliferative peak of hepatocytes. We demonstrated that OPN depletion leads to impaired liver recruitment of macrophages and neutrophils, blunts the activation and inhibits IL-6 production in Kupffer cells, and consequently attenuates the activation of the Stat3 pathway in hepatocytes. Therefore, OPN is an additional molecule that may promote the initiation of liver regeneration.
Loss of OPN impairs the recruitment of macrophages and neutrophils into the regenerating liver. (A) Liver leukocytes were isolated and analyzed at 0 and 3 hours post-PHx, and the number of liver leukocytes (Neutrophil: Gr-1high CD11b+; Macrophage: CD11bint F4/80+) was calculated by multiplying the percentage of each population determined by the FACS data by the total number of leukocytes per liver (n = 4). The data are shown as the mean ± SEM. *P<0.05. Immunohistochemistry for (B) MPO and (C) F4/80 were performed in mouse liver sections at 3, 6, and 12 hours post-PHx (400× magnification). Quantitative analysis was performed (n = 3-5). The data are shown as the mean ± SEM. *P<0.05, **P<0.01. (D) MPO and (E) F4/80 relative expression levels were analyzed in livers after PHx (n = 3-4). The data are shown as the mean ± SEM. *P<0.05. HPF: high power field; RFC: relative fold change.
OPN facilitates IL-6 release in the regenerating liver and isolated Kupffer cells. (A) Serum IL-6 levels were determined by ELISA after PHx (n = 4-6). The data are shown as the mean ± SEM. *P<0.05. (B) IL-6 mRNA expression in the liver was analyzed by qPCR (n = 3-5). The data are shown as the mean ± SEM. *P<0.05. (C) Kupffer cells were isolated from WT mouse livers and treated with 100 nM rmOPN. Cell culture supernatants were subjected to ELISA for IL-6 (n = 3). The data are shown as the mean ± SEM. (D) Cell lysates were collected for quantitative analysis of IL-6 mRNA (n = 3). The data are shown as the mean ± SEM. *P<0.05. RFC: relative fold change.
OPN facilitates IL-6 production in Kupffer cells and promotes liver regeneration via activation of the Stat3 pathway. (A) Kupffer cells, hepatic stellate cells and hepatocytes were isolated from WT and OPN-/- mice after PHx and collected for quantitative analysis of IL-6 mRNA (n = 3-6). The data are shown as the mean ± SEM. **P<0.01. (B) Phosphorylation of Stat3 in regenerating livers was analyzed by western blot. Densitometric analysis of immunoblots is shown (n = 3). The data are shown as the mean ± SEM. *P<0.05, **P<0.01. (C) mRNA expression of c-fos, c-myc, and c-jun was analyzed by qPCR (n = 3-6). The data are shown as the mean ± SEM. *P<0.05, **P<0.01. RFC: relative fold change.
Consistent with a previous finding , we observed that OPN is exclusively expressed in BECs in normal livers. Furthermore, after PHx, the majority of induced OPN was located in the areas of the bile ductal epithelium. Here, we used CK19 as the specific marker for BECs, as shown in Fig. 1D. It has been reported that oval cells also stain positive for CK19, but they are usually absent in a normal liver and are generally activated 5-10 days post-PHx [17, 18]. Hence, we believe that the CK19-positive cells observed at 3 hours post-PHx are mainly BECs. Therefore, we concluded that BECs are primarily responsible for OPN production after PHx.
Compared to the control group, we observed a delay of hepatocyte proliferative peak after PHx in mice deficient in OPN; however, we did not observe a proliferative advantage in hepatocytes treated with recombinant mouse OPN (rmOPN) in vitro, suggesting that OPN does not directly stimulate hepatocyte proliferation. Interestingly, a recent study reported that recombinant human OPN (rhOPN) decreased primary hepatocyte proliferation but did not alter cell viability . Given that the recombinant mouse OPN shares only 64% amino acid sequence identity with human OPN, we suspect that this discrepancy might be due to different types of rOPN. Further studies examining whether OPN can directly affect hepatocyte proliferation are warranted.
In the present study, we demonstrate that ablation of OPN impaires recruitment of macrophages and neutrophils into the liver during the priming stage following PHx, which may contribute to the reduced IL-6 levels observed in OPN-/- mice. We also detected that WT Kupffer cells produce more IL-6 in vivo at 3 hours after PHx compared to OPN-/- Kupffer cells, suggesting that, in addition to the OPN-mediated hepatic infiltration of macrophages, OPN may also enhance IL-6 production in Kupffer cells, either directly or indirectly. Notably, in vitro direct incubation of Kupffer cells with rmOPN slightly induced IL-6 production, which raises an interesting question: how does OPN modulate the remarkable IL-6 induction in Kupffer cells after PHx? We then examined LPS levels in the liver and in portal circulation. Significantly higher LPS levels were detected in both in WT compared to OPN-/- mice, suggesting that after PHx, OPN may affect LPS release into the portal vein and subsequently modulate IL-6 induction in Kupffer cells. Intriguingly, in an alcohol-induced liver injury mouse model, a recent study found that OPN blocks the gut-derived LPS into the liver by binding to LPS . Therefore, we propose that OPN might affect the stimulation of LPS release in different experimental models.
The IL-6/Stat3 pathway may affect the induction of nearly 36 percents of the immediate early genes involved in the regeneration of the liver [23, 34]. Among them, c-fos, c-myc and c-jun play pivotal roles in liver regeneration. c-fos is positively transactivated by the Stat DNA binding element; c-myc transcriptionally regulates cyclin D1, the best marker for the G1-to-S transition in the regenerating liver; and c-jun acts as a positive factor in cell growth . In OPN-/- mice, the expression of all three of these genes was repressed due to the reduced activation of the Stat3 pathway, which is the key process that activates hepatocytes to enter the mitosis .
In summary, to the best of our knowledge, we have for the first time demonstrated the positive regulatory role of OPN in liver regeneration. Deletion of OPN delays the peak of hepatocyte proliferation but does not impair overall liver regeneration. This delay in proliferation in OPN-/- mice post-PHx seems to be results of a decrease in IL-6 production, which is attributed to the impaired hepatic recruitment of macrophages and the reduced LPS release to portal circulation observed in OPN-/- mice after PHx. Thus, our study sheds additional light on the molecular mechanisms that regulate the process of liver regeneration.
OPN: osteopontin; IL: interleukin; Stat3: signal transducer and transcription 3; TNFα: tumor necrosis factor α; IL-6R: IL-6 receptor; IL-22R: IL-22 receptor; NF-κB: nuclear factor κB; Spp1: secreted phosphoprotein 1; Eta-1: early T-lymphocyte activation 1; IFNγ: interferon γ; PHx: partial hepatectomy; BrdU: 5-Bromo-2'-deoxyuridine; ALT: alanine transaminase; AST: alanine transaminase; PCNA: proliferating cell nuclear antigen; MPO: myeloperoxidase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HSC: hepatic stellate cell; BEC: biliary epithelial cell; LPS: lipopolysaccharide; HGF: hepatocyte growth factor; CK19: cytokeratin 19; RFC: relative fold change; rmOPN: recombinant mouse OPN; rhOPN: recombinant human OPN; SEM: standard error of mean; HPF: high power field.
We thank Dr. Bin Gao from National Institutes of Health, Bethesda, MD for editing the manuscript. This work was supported by the National Natural Science Foundation of China (31300742 to X Kong, 81372233 to H. Wu, 8147224 to Q Xia, 81130038 and 81372189 to WQ Gao), and the Shanghai Education Committee (Eastern Scholar Program) to X Kong, the Chinese Ministry of Science and Technology (2012CB966800 to WQ Gao), the Shanghai Health Bureau Key Joint Efforts Foundation (2013ZYJB001) to Q Xia, the Shanghai Health Bureau Key Discipline and Specialty Foundation and the KC Wong Foundation to WQ Gao.
All authors have read the manuscript and declared that there is no conflict of interests.


Fenerbache will look to continue from where they stopped as they hope to move up to a decent position on the Super Lig table.
Fenerbache spent most part of the first stanza in the drop zone, however the transfer window has helped the team to boost their squad.
The super Lig full of Premier League ex-stars are all making waves in their different clubs.
Kayserispor hasn't been convincing lately, having two draws in the last two matches isn't a good show for a team close a to the drop zone.
Fenerbache won convincingly against Goztepe in their last Super Lig matche courtsey of goals from Ayew and Victor Moses.
Fenerbache have the upper hand in the encounter due to their performance in recent matches.
Fenerbache went into the transfer window in search of better players that could change their situation in the league.
Kayserispor on the other hand hasn't played particularly well, but a series of draws at home and away in recent matches.


Swiss skier Martina Schild, who collected silver in the downhill at the 2006 Turin Olympics, has decided to retire due to injury, the Swiss ski federation announced on Thursday.
The 31-year-old bows out having missed all of last season with a dislocated hernia.
Schild takes leave of the slopes with three World Cup podium finishes to her name, her lone victory coming in the super-G at Lake Louise in 2007.
At the Turin Games, she sprang a surprise when beating all bar Austria's Michaela Dorfmeister in the downhill.
Schild is the grand-daughter of Hedy Kaufmann-Schlunegger, winner of the inaugural women's downhill gold medal at the 1948 Olympics staged in St Moritz.


text:
Reprezentacja Estonii w hokeju na lodzie mężczyzn – kadra Estonii w hokeju na lodzie mężczyzn.

Trenerzy
Trenerem kadry był Dmitrij Miedwiediew. W sezonie 2013/2014 kadrę prowadził Fin Sakari Pietilä, po czym następcą został jego rodak Jussi Tupamäki.

Mecze reprezentacji Estonii (1937-1940)
Zestawienie oficjalnych międzynarodowych meczów reprezentacji Estonii w hokeju na lodzie mężczyzn.

Udział w mistrzostwach świata
 1993: niesklasyfikowana (2. miejsce w Grupie C2 kwalifikacje)
 1994: 28. miejsce (1. miejsce w Grupie C2)
 1995: 24. miejsce (4. miejsce w Grupie C1)
 1996: 25. miejsce (5. miejsce w Grupie C)
 1997: 23. miejsce (3. miejsce w Grupie C - awans)
 1998: 19. miejsce (3. miejsce w Grupie B)
 1999: 22. miejsce (6. miejsce w Grupie B)
 2000: 22. miejsce (6. miejsce w Grupie B)
 2001: 28. miejsce (6. miejsce w I dywizji - spadek)
 2002: 29. miejsce (1. miejsce w II dywizji - awans)
 2003: 22. miejsce (3. miejsce w I dywizji)
 2004: 22. miejsce (4. miejsce w I dywizji)
 2005: 23. miejsce (4. miejsce w I dywizji)
 2006: 24. miejsce (4. miejsce w I dywizji)
 2007: 23. miejsce (4. miejsce w I dywizji)
 2008: 27. miejsce (6. miejsce w I dywizji - spadek)
 2009: 31. miejsce (2. miejsce w II dywizji)
 2010: 29. miejsce (1. miejsce w II dywizji - awans)
 2011: 27. miejsce (6. miejsce w I dywizji - spadek)
 2012: 29. miejsce (1. miejsce w II dywizji Grupie A - awans)
 2013: 28. miejsce (6. miejsce w I dywizji Grupie B - spadek)
 2014: 29. miejsce (1. miejsce w II dywizji Grupie A - awans)
 2015: 27. miejsce (5. miejsce w I dywizji Grupie B)
 2016: 27. miejsce (5. miejsce w I dywizji Grupie B)
 2017: 26. miejsce (4. w Dywizji I, Grupie B)
 2018: 25. miejsce (3. w Dywizji I, Grupie B)
 2019: 26. miejsce (4. w Dywizji I, Grupie B)

Przypisy

Linki zewnętrzne
 Oficjalna strona federacji
 Informacje na stronie IIHF

Estonia
Hokej na lodzie w Estonii
Estońskie reprezentacje narodowe

redpajama_set_name: RedPajamaWikipedia


text:
About V I
Breakdown Chapter Four
Charms–Or Something
As I walked along Chicago Avenue, a couple coming out of a bar tried to offer me a dollar for a cup of coffee. Their gift made me realize just what a bizarre vision I must present—in my running shoes and bedraggled evening gown I was an avatar for homelessness.
I'd started the evening looking like I belonged in a limo, or at least in the grand ballroom at the Valhalla Hotel, which is where I'd been headed. I've never been fond of big glitzy events, and you go to the Valhalla only if glitz is your middle name. I especially wasn't fond of them when they celebrate the life and work of people I despise: in this case, Wade Lawlor.
If you don't know Lawlor, it's because you get all your news from microform copies of the Chicago Daily News. Local boy made, well, "good," would be putting a values spin on it. Local boy made national superstar was more like it.
Although I tried never to watch the show, you can't live in Chicago and not know Lawlor's face—it's on the sides of buses, on billboards, on the back of the Herald-Star. GEN, the Global Entertainment Network, whose lead cable news show Lawlor hosts, often features him on its billboard along the Kennedy Expressway.
Lawlor's signature is a blue-checked work shirt, open at the throat to show he's a working man who scorns the suit and tie of an effete liberal journalist. His thick black hair is artfully tousled on-air: America's in danger, I don't have time to comb my hair!
For his anniversary carnival, Lawlor incongruously wore his checked shirt with a dinner jacket, a modern one with square pockets sewn to the jacket front. An American flag picked out in jewels was on the lapel. It had a fancy little ear of corn on top of it, as if to point out that he could afford diamonds and rubies, but he was basically a Midwest hick at heart.
Lawlor was working the room with one of those top-grossing stars whose name and face you keep seeing in Us and People. My red evening dress is a backless, ankle-length number, but the star, whose smile seemed epoxied in place, made me feel overdressed. When Lawlor came over to where I was standing with Murray Ryerson, I tried, discreetly, to see how his date kept her breasts from tumbling out of the front of her dress, since it opened all the way to her waist. More epoxy, I decided, keeping a glass in my right hand and food in my left so that I wouldn't have to touch Lawlor.
"Hey, Ryerson, thanks for showing up." Lawlor's eyes scanned the room behind us, looking for people more worth his attention.
"Wouldn't miss it," Murray said with unnecessary heartiness.
Lawlor smirked. "And who's the talent?"
"V. I. Warshawski," I said.
"I haven't seen you before. Out of town?"
"Totally local," I assured him. "Steel City. And you?"
"What's 'Steal City'? The Chicago motto?"
"Very clever, Mr. Lawlor. I'll have to put that in my blog, how clever you are, and what a thrill to meet you, and so on."
I kept my voice languid, trying, for Murray's sake, to keep the venom I felt out of it. Even so, Lawlor's lips tightened and his eyes narrowed. He put his hand on the star's elbow and started to guide her away, but she stayed put. Perhaps she didn't like him any better than I did; perhaps it was her publicist's idea that she be seen with him on the entertainment sites.
"Are you with GEN?" she asked.
"I'm a private investigator," I said. "Murray Ryerson and I have worked together on a number of stories."
Lawlor eyed me in a way that made me long to take his ribs apart. "She your legwoman, Ryerson? Why'd the network give you the one with the body and me the ugliest guy in Chicago?"
"I guess our looks match our results," I said.
Lawlor frowned; the veneer of charm vanished to expose a startling rage. "What's that supposed to mean?"
Murray knocked my arm hard enough that the wine sloshed over the rim of the glass. We all exclaimed at the mishap, and the star let Lawlor lead her on.
"Why the fuck did you have to say that?" Murray demanded.
"It was just banter, Murray. I didn't know he was sacred and that you're not allowed to answer back to his gibes. Is it critical for your career for me to find him and apologize?"
"No, no, don't!" Murray said. "Your apology might involve black eyes and stuff that would really end my career."
"And that flag pin—does he have that glued to every garment he owns?" I fumed. "What's with the ear of corn? Is he showing that he's the corniest man in America?"
"Where have you been since campaign season started, Vic? That's Helen Kendrick's signature—U.S. flag with corn from the heartland. Ethanol is a big chunk of her husband's family fortune, you know that. And Lawlor is her number-one booster."
Kendrick was running for Senate. She thought the last time America had been a great country was the day before Lincoln signed the Emancipation Proclamation, so it was no surprise that Lawlor backed her campaign.
Various other media and entertainment celebrities drifted by. If I did keep a blog, I'd have written up the number of national figures who felt their careers required them to get freshly Botoxed and epoxied, and show up in little numbers by Chloé or Vera Wang. I didn't care about spotting stars of GEN's reality show All-American Hero. What staggered me were the senators and even Supreme Court justices who'd flown in from Washington to see and be seen. That told a sobering tale of how influential Lawlor's voice was on America's political scene.
A few minutes later, Harold Weekes, head of GEN's news division, ambled by. Even though I thought he was the slime on the pond, I smiled, said little or nothing, and even let him leer at my cleavage.
"Keep up the good work, Ryerson! 'Chicago Beat' matters to us at Global One, you know."
I couldn't help rolling my eyes at that—what a name for the ugliest chunk of glass and steel to go up in Chicago since Trump Tower broke ground.
"I'm happy to hear that, Harold," Murray said, with an effusiveness that made me wince. "I wanted you to meet Vic here. V. I. Warshawski. She's one of Chicago's most skilled criminal investigators."
Weekes's brows went up. "Expecting to find murder here?"
"Nope," I said. "Just the usual graft and corruption, nothing special."
"Vic has done background work on a lot of my stories," Murray said hastily. "Last winter's exposé on war profiteering, for instance."
Weekes frowned. "I know you thought you had a big scoop there, Ryerson, but it's always been true that war creates opportunities for the alert."
I grinned insanely, the little woman ecstatic to be in the presence of power. "For the alert opportunist, I suppose. Other people just have the chance to get their heads blown off."
There was an uncomfortable silence for a beat, and then Weekes laughed. A smiling woman in a silky red dress, she could be given the benefit of the doubt.
Murray plowed ahead doggedly. "You know the series I'm working on, 'Madness in the Midwest,' on the mentally ill, from the streets to state hospitals' forensic wings—Vic could add a lot of depth to the series."
Weekes patted Murray's arm. "We'll certainly keep that in mind, Ryerson. If your friend has investigative experience we can probably find a role for her."
Like Lawlor a few minutes earlier, Weekes's eyes were glazed over. Talking to Murray and his friend was his idea of purgatory. I couldn't really blame him—the feeling was completely mutual.
The governor of Wisconsin came along and tapped Weekes's arm. The news king moved on.
"Murray, is that why you invited me to this horror show? To help you with some story about mentally ill criminals?" I demanded. "Why didn't you tell me you had an agenda?"
"You put up such a song and dance about coming at all, I didn't feel I could go into it with you," he blazed back.
That much was true. When Murray called last week, asking me to be his date to tonight's celebration of Lawlor's tenth anniversary as GEN's star, I'd said no without thinking.
"I hate Wade Lawlor," I protested. "I hate his politics, I hate his molassied voice, and I hate his pretense of being a working-class boy. That fake work shirt makes me throw up every time I drive up the Kennedy. I bet the closest he ever got to a day's hard labor was paying a neighbor to mow his mother's lawn when he was a kid."
"You'd lose," Murray said. "He comes from some kind of broken home. I've seen him weep on camera, over how his dad ran off and left him and his sister to fend for themselves. Success hasn't just gone to Wade's head—it's made him vindictive. My job ain't so secure that I can dis the network's golden goose. And I don't want to go alone."
"What about all those blond twentysomethings you flaunt any time I see you in public?" I snapped.
"Are you jealous?"
"More disgusted. Why can't you act your age?"
"That's what I'm trying to do here, and you're not helping. You can rehabilitate me, turn me into a boomer who's not afraid to show his age," he wheedled.
"If I could rehabilitate you, Murray, it would be to turn you back into the journalist who won a Pulitzer for White Crime/Black Convict."
As soon as the words were out of my mouth, I'd wished them back. Murray had done a spectacular series for the Herald-Star on the white youths who bought their coke and meth on the black South and West Sides but who were seldom accused of drug crimes. He'd followed four kids—two black, two white—from the drug scene for a year. At the end of the year, the two white guys were setting off for the east coast to college at Haverford and Princeton; one of the black kids had been sentenced to fifteen years for possession, while the other was dead.
The year after Murray won his Pulitzer, the Global Entertainment Network bought the Star, along with several hundred other papers. Harold Weekes acquired a minor Hollywood studio for its cable potential, moved the company from the outer reaches of LA to Wacker Drive, gutted the reporting staff at Global's papers, and hired Wade Lawlor to disseminate rumors, innuendo, and outright lies, under the catchy title "Wade's World."
"Wade's World" trumpeted the claim that Obama had ordered police to collect Bibles from kids on their way to school. Wade signed on with a group disputing the president's citizenship.
It wasn't Murray's fault that his new owners preferred video to print. It wasn't his fault that he had to scramble to keep a job, so I didn't hold it against him that he anchored a weekly TV show on GEN. In "Chicago Beat," he reported on everything from politics to the arts, but most of his shows were devoted to sensational crimes, since that's what draws a crowd.
Despite Harold Weekes's hearty assurance that everyone in GEN's headquarters loved "Chicago Beat," Murray's show aired once a week in Illinois and Indiana, although Wisconsin and Michigan affiliates sometimes picked it up. "Wade's World" was shown four times daily in every city, village, and farm in America.
I didn't believe Murray's career hung by the thread he kept claiming. But he was working in a poisonous environment. Lawlor was reported to pull in twenty million a year just from GEN, while his endorsement contracts probably tripled that figure. Other GEN cable stars made seven-figure salaries; in a milieu where the chief operating officer dismissed print journalism as "turning back the clock to the era of illuminated manuscripts," no matter what Murray earned, he was bound to feel insecure.
I felt ashamed of rubbing Murray's face in his troubles; I said I'd go to Lawlor's event at the Valhalla with him. And I'd regretted it the minute I walked in the door. After the encounter with Weekes in the Valhalla ballroom I was furious.
"You could have warned me before I met your boss that you wanted my help with a story. I could have jumped in with some intelligent backup, instead of which he dismissed me as your girlfriend."
Murray looked sheepish. "I just couldn't find a way to propose it to you, and then Weekes popped up, and I wanted to get the idea back in front of him. You saw the first piece, looking at the returning Iraq/Afghan vets who've become homeless."
"Yes, yes, I did. You did a great job with that. I didn't know it was the start of a series, though."
"It wasn't," he fumed. "I had the whole series started—I was going out to one of the state mental hospitals to look at murderers found not guilty by mental defect, I had one on the advanced practice nurses who do most of the hands-on medical care of the mentally ill homeless—I had nine shows lined up, and I had my own producer's blessing, and then, right after the one on the vets ran, Wade Lawlor stuck in an oar at the huddle, said it was banal and a resource-eater, and Weekes axed the whole series."
"I can't possibly persuade Harold Weekes to listen to you instead of Lawlor," I protested.
"No, but I was hoping you could come on board as the resident expert on evaluating criminal evidence for the segment on people found not guilty by reason of mental defect. I've tried pitching that again as a single episode; I put together a list of five people who've been held at Ruhetal or Elgin for more than twenty years, but Lawlor keeps shooting down the idea, and I never get time alone with Weekes. I was hoping if he saw you, I don't know—"
"Even though I'm not a blond twentysomething, that my sparkling gray eyes and flawless skin would captivate him."
Murray grimaced. "You have a way of putting things in the worst possible light, but, yeah, something like that."
It was then, as the noise level in the ballroom had passed the dangerous decibel mark, Petra's call came, begging me for help. I told Murray I had a client in trouble, refused to give him details, and fled. It took a good fifteen minutes for the Valhalla valet to fetch my car. By the time I got to Mount Moriah, the girls had a substantial head start on me.
Copyright © 2019 By Sara Paretsky // Privacy Policy // Contact // Site Design By Lisa Hazen Design & Editorial

redpajama_set_name: RedPajamaCommonCrawl


You hear someone speak and immediately guess that she is from Australia. Why? Because her accent reminds you of other Australians you have met. Or you try a new restaurant expecting to like it because a friend with good taste recommended it. Both of these are examples of decisions based on experience. When faced with new situations, people are guided by memories of similar situations that they have experienced in the past. That is the basis for the data mining techniques introduced in this chapter.
Nearest-neighbor techniques are based on the concept of similarity. Memory-based reasoning (MBR) results are based on analogous situations in the past — much like deciding that a new friend is Australian based on past examples of Australian accents. Collaborative filtering (also called social information filtering) adds more information, using not just the similarities among neighbors, but also their preferences. The restaurant recommendation is an example of collaborative filtering.
Central to all these techniques is the idea of similarity. What really makes situations in the past similar to a new situation? Along with finding the similar records, there is the challenge of combining the information from the neighbors. These are the two key concepts for nearest-neighbor approaches.


text:
Shiites flee enclave in Pakistan after Taliban lay siege
By Jane Perlez and Pir Zubair Shah, International Herald Tribune, July 27, 2008
PESHAWAR, Pakistan: It was once known as the Parrot's Beak, a strategic jut of Pakistan that the U.S.-backed mujahedeen used to carry out raids on the Russians just over the border into Afghanistan. That was during the Cold War.
Now the area, around the town of Parachinar, is near the center of the new kind of struggle. The Taliban have inflamed and exploited a long-running sectarian conflict that has left the town under siege.
The Taliban, which have solidified control across the Pakistani tribal zone and are seeking new staging grounds for attacking U.S. soldiers in Afghanistan, have sided with fellow Sunni Muslims against an enclave of Shiites settled in Parachinar for centuries. The population of about 55,000 is short of food. The fruit crop is rotting, residents say, and the cost of a 30-kilogram, or 65-pound, bag of flour has skyrocketed to $100.
And, in a mini-conflict that yet again demonstrates the growing influence of the Taliban and the Pakistani government's lack of control over this sensitive border area, young and old, wounded and able-bodied, have become refugees in their own land.
Thousands of displaced Shiites from Parachinar are scattered among relatives in Peshawar, capital of North-West Frontier Province, which abuts the tribal areas, and in hotels and shelters where images of Iranian religious leaders decorate the halls.
For Complete story, click here


The Calvert County Election Board will conduct a manual ballot audit of the 2018 General Election on Tuesday, February 12, 2019 starting at 10 am in Conference Room 1, 30 Duke St, Prince Frederick*.
- If schools are cancelled, we will move the audit to Wednesday, February 13.
- If schools and/or the Calvert County government are on a 2-hour delay, we will start the audit at 11 am on Tuesday.
The Calvert County Election Board registers any eligible person who wishes to vote and is responsible for all registrations for county, state, and national elections, as well as the registrations for the municipalities of the county. This office also helps persons seeking elective office with candidate filing and maintains electoral information.
The staff and board members of the Calvert County Election Board are committed to providing the highest level of customer service to all voters and prospective voters in Calvert County. We are dedicated to ensuring that all voters can vote in private, in a trouble-free environment, and be assured that their vote will count.
Please visit the Maryland State Board of Elections website for complete information.
Early voting occurs over an 8-day period before Election Day. The Early Voting Center for Calvert County registered voters is located in the lower level conference rooms of the Community Resources Building (same as office). For more information, please visit the Maryland State Board of Elections website. For important voting dates, please see our calendar.
The Calvert County Board of Election Members meet monthly on the fourth Tuesday at 2:30 p.m. in Conference Room 1, near the Election Board Office.
Do I have to register to vote before each election?
What is the last day to register to vote or make any changes to my registration?


Thank you to all our partners & sponsors!
We invite you to be part of an innovative approach to connecting educators in the fields of literacy and digital technology by becoming a conference sponsor. The 4T Virtual Conference on Digital Writing (Teachers Teaching Teachers about Technology) is a unique opportunity for K-16 educators and administrators to revolutionize how instruction and professional development is conducted. This is a one of a kind conference that makes possible the development of technical and pedagogical skills required for effective teaching of digital writing. It is a satellite of the 4T Virtual Conference, which is sponsored by the University of Michigan's School of Education.
The 4T Virtual Conferences fundraise together and reach over one thousand participants from around the world. We expect a very high turnout for the Digital Writing Conference because of its affiliation with both UM's School of Education, a large Intermediate School District, and National Writing Project sites in Michigan. Participants will be digitally active literacy educators who will tweet heavily about the conference and the products they experience during the event.
By becoming a conference sponsor for both 4T conferences, you open the door for increased exposure of your company and products. Don't miss out!
Company logo and a link to your website on the homepage of the 4T and 4TDW Conference website.
Company logo and a link to your website on the opening slide in every 4T and 4TDW conference session.
Company logo and a link to your website will be included in our twice-monthly newsletter between January, 2016-October, 2016.
Company logo and a link to your website with featuredplacement on the homepage of the 4T and 4TDW Conference website PLUS a link to a video demonstration of your tool or company.
Company logo and a link to your website with featuredplacement on the opening slide in every 4T and 4TDW conference session PLUS the moderator at each conference session will verbally acknowledge your company.
Company logo and a link to your website will be featured in our twice-monthly newsletter between January, 2016-October, 2016.


MANILA: Philippine President Rodrigo Duterte has urged China to "temper" its behaviour in the South China Sea in a rare criticism of the Asian superpower over its programme of island-building in disputed waters.
China has alarmed and angered its neighbours by claiming dominion over most of the South China Sea and building a string of artificial islands and military air bases.
But the outspoken Duterte – keen to court trade and investment from Beijing – has mostly withheld criticism.
In a change of tone, Duterte said in a speech late on Tuesday to business entrepreneurs that China had no right to claim airspace above man-made islands.
Philippine officials have claimed military pilots are repeatedly warned off by Beijing as their planes approach Philippine-held Thitu island, which lies beside a Chinese air base built on top of Subi Reef.
"You cannot create an island. It's man-made and you say that the air above this artificial island is yours," Duterte said, according to a transcript released by the presidential palace yesterday.
"That is wrong because those waters are what (one) would consider international sea. And the right of innocent passage is guaranteed," said Duterte, who did not refer to any specific incident.
He added that he did not want to "quarrel" with China.
The comments follow allegations in May of Chinese harassment of Filipino troops at another South China Sea garrison.


text:
NEW ARTIST... 88 LO
Published: May 3, 2019 by Shu Banga Bloggs Filed under: HipHop & Rap - 64 Views
Introducing amazing new artist, song-writer and C.E.O., 88 LO. He is far from your average rapper. A triple-threat, he raps, produces and sings his own hooks. He is also C.E.O. of his own label, 88 MUSIC GROUP, LLC. His innate ability to think beyond his circumstances and great business mind definitely put him ahead of the curve.
From very early on in his life, music was a source of healing and seemed to ease the blows life dealt him. Growing up in Jamaica he was exposed to the violence and drug wars soundtracked by the island vibes of Bob Marley, Beres Hammond, Peter Tosh and Jimmy Cliff. At that time the DJ's were gangsters and they ran the gangs, dance halls and the music scene. This fascinated LO. "I remember seeing Supa Cat, Louie Rankin, Capleton and Tiger come through the dance halls dapper and drippin' with jewels. They were superstars, they were Dons. Music was sort of an escape for the real gangsters from the killing, drugs and gangs that plagued our towns in Jamaica. You could feel the pain in the beats and rhythms of their songs. They inspired me to become an artist."
Born Jamaal in Kingston Jamaica West Indies, LO came to the United States at the age of 15 yrs old. He grew up in the Bronx, N.Y. in a neighborhood plagued with drugs, crime and gangs, very much like Kingston. He quickly fell in with the wrong crowd and eventually, he found himself incarcerated and facing 25 years to life. At that moment he decided he needed to change the direction his life was headed in. He got on his knees and prayed to God with all his might for forgiveness and promised to walk a straight path. His prayers were answered, he was found not guilty and released from prison. Now it was time to make good on the promises he made to God.
88 LO began his transition from the street life to an artist and businessman investing his time, money and full energy into studio sessions and creating his label. He and his friends started their own group called 88. Each member had a different skill and talent to contribute to the group. Some sang, others rapped and/or made beats. In 2017, 88 LO, 88 GOLDIE, 88 CedDean and 88 K-Dogg released their first track, "TrapHouse" with a visual for YouTube. It received close to 500 views! This was just the beginning...
The hard work payed off when DJ K-SLAY caught wind of the single, "TrapHouse". He was quite impressed with the song, in particular, 88 LO's verse. He reached out to LO and they began to build a working relationship. In late summer 2018 LO released his first solo single, "Betrayed", to rave reviews! The video has garnered over 30, 000 views on YouTube to date. Six months later he dropped his first official project, the "Millionaire Dreams and Billionaire Nightmares" mixtape hosted by DJ K-SLAY.
His style is authentic and doesn't mimic any sound you hear on the airwaves. His sound infuses DanceHall and Reggae with gritty Hip-Hop. His musical roots in Jamaica are evident and provide the right spices to the ingredients for this mouthwatering dish! "Everything about me is authentic. The shit I rap and sing about is all real and inspired by my life experiences. I am a Jamaican-born emcee who raps New York-style and I sing my own verses. That makes me unique and amazing! Originality is missing from the music right now. I have always been a leader and most artists are followers nowadays. I've come to restore some order, know what I mean?"
88 LO's latest project, yet to be titled, is slated to be released summer of 2019. Seven straight bangers with guest features from Dave East and Tory Lanez. New York is sounding like New York again!
INSTAGRAM: 88_LO_
https://youtu.be/T-ypwnrx0kQ
Shu Banga Bloggs
Free Lance Writer - Journalist
Tags: 88 LO;
Hot new artist LiL RAFE!
SouthSide Jamaica, Queens has a diamond in the ruff by the name of Tyise Troy Garrett aka LiL RAFE. RAFE was introduced to...
New Artist: Big Boss Mulaa
Introducing new artist Big Boss Mulaa from SouthSide Jamaica, Queens. Mulaa started his career on the opposite side of the...
new artist...JP CALI SMOOV
MeetJP Cali Smoovmaster lyricist and aspiring actor from Los Angeles, California. JPhas been making music his entire life ...


This portion of Gate 61/2 Road is poorly maintained. It has ruts and potholes that are getting worse due to frequent and increasing car traffic from out of town bikers who park there while riding on the bike path. Residents who live along the road are risking potential damage to their cars each time they drive on it.


text:
Exclusive: "Mifune: The Last Samurai" director, Steven Okazaki said "you realize how special the Kurosawa movies are."
By: Izumi Hasegawa December 5, 2016
Toshiro Mifune is the one of the greatest Japanese actors in history, appearing in over 170 films. Although I too am Japanese, I did not know the details of his illustrious career and why he was labeled as one of the greatest.
Then I saw the film, Mifune: The Last Samurai, and not only do I now completely understand why, I fully agree with the statement .
Mifune is a documentary directed by Japanese American director, Steven Okazaki. Although Okazaki was born in Venice Beach, he has been to Japan enough to be very familiar with the culture and the film history of Japan, so the film delivers a very authentic view of the Japanese film industry. I was able to connect with Steven by phone and he shares what the process for Mifune: The Last Samurai and the history of Japanese films.
Q: This film is so beautifully shot. How did you go about that?
It's difficult to interview Japanese people in their home since they're always self-conscious about letting you in. We found this great place right outside of Tokyo called The Edo Architectural Museum Park, and they preserved buildings from all over Japan. They just lift them up and bring them to the park. We used different locations in this park outside of Tokyo, so that made it easier.
Q: Why did you decide to make the Mifune documentary?
Well I was originally interested making a documentary on the history of Chanbara (sword fighting) films. I was really interested in trying to see the old Chanbara silent films. I wanted to see the history of 100 years of Chanbara movies. That's very difficult and so many of the movies have disappeared. You would have to work with different movie studios in Japan, and that's very difficult (to work with different studios). I heard that a producer wanted to make a film about Mifune, and I got very excited. I said, "Please let me do it." I met with them, and they said, "Okay let's do it."
Q: You have so much archival footage; I didn't know a Chanbara silent film existed.
There are not too many, they're precious. I think the saddest thing is how many great films no longer exist, especially the films in Japan before WWII. Some films were destroyed, or people didn't care for them. Very few of the films survive. Some of the films pop up in foreign countries, and they found them there.
It's a very interesting thing. People think of Japan that is a culture those years surpassed. I don't think that's true. When you have a culture that's preserved in paper and wood, it's very hard to preserve it. There's been pieces of Japanese history, including film history that no longer exists. The few old films that exist are very precious, and many of them are in terrible shape.
Q: What was your biggest concern shooting this film?
My big worry was Mifune was born in 1920 and you had this wonderful time where everyone was roughly around the same age. It was a very exciting time. After the war you had new people who wanted to be actors or directors or cameramen. They were all kind of the same age. My big worry for the film was finding enough people that knew Mifune who were still alive. The average age of the people was between 80 or 85. They were all very "Genki" (energetic in Japanese) and when you mentioned Mifune, their eyes lit up and loved to talk about him.
Q: How did you approach the scheduling for these famous people in the documentary including Steven Spielberg and Martin Scorsese?
Spielberg and Scorsese, we did have to wait for the schedule and it was very quick. They would say, "We have one hour" and we would wait for them. When we asked them, they immediately said yes, but it took a while to get to them. When we finally did (get to them) they were very enthusiastic. They had only admiration for Kurosawa.
Q: You chose Keanu Reeves for narration which is an amazing choice, but I am curious why you went with him since his voice isn't as emotionally expressive like Samurai's was.
Just to edit with the film, I used my voice. Although it's very painful for me to listen to my own voice, other people said, "that's okay, we can use that." We thought it would've been interesting to have more English language in the film. We thought about many actors but some of them have very distinctive style of sounding very New York, or sounding very Californian.
I was also worried about their pronunciation of Japanese names and movies. We spent too much time on that I just thought that something about Keanu Reeves sort of fit Mifune. He doesn't act like a big movie star, and he's not surrounded by agents and managers. He just shows up by himself on his motorcycle. He loves making movies but he doesn't take himself too seriously and just seems like more like the regular person Mifune was. We thought that might work and so one of the producers was interpreter for him in his movie "Man of Tai chi". He asked Keanu and he said sure.
Q: While I discovered why Mifune he was such a terrific actor and that he actually got his start as a baby model for his father's photo studio. What did you discover about Mifune while working on this film?
You really take movies for granted. You go, "oh that's fantastic." I don't know what the movie of the moment is. When you watch the movies and work on a film like this you realize how special the Kurosawa movies are. They're not typical of Japanese movies made of the same time. The budgets and schedules were much more difficult at the time. Kurosawa really had this strong ego and determination to make something that people would really notice something different. If you look at the other movies made at the time, there's nothing that really can compare with the kind of care in Mifune (that) Kurosawa put into it.
That's the one thing; the actress Tsukasa (Yoko) says when Kurosawa and Mifune stopped making films. It was an end of an era for all of us, not just for Kurosawa and Mifune. We had to come back down to earth in a way. I think the specialness of that time and appreciation of it. It doesn't happen in movie making all around the world. That really was a special time in Japan.
Mifune: The Last Samurai is playing on theaters now.
Interview by Izumi Hasegawa – @HNW_Izumi
Edited by: Jody Taylor – @RealJodyTaylor
Follow Us: What's Up Hollywood at @WhatsUpHWood
Hollywood Event Calendar
Exclusive!!! Carmen and Lupita Andrade from "Conjoined Twins: Inseparable" premieres on Thursday, July 16th, 10pm on TLC
June 22th is Chris Pratt's Birthday. Happy Birthday, Chris! Let's revisit his interview!
June 13th is Chris Evans's Birthday. Happy Birthday, Chris! Let's revisit his interview!
June 9th is Natalie Portman's Birthday. Happy Birthday, Natalie! Let's revisit her interview!
June 9th is Johnny Depp's Birthday. Happy Birthday, Johnny! Let's revisit his interview!
June 7th is Liam Neeson's Birthday. Happy Birthday, Liam! Let's revisit his interview!
Hollywood News Wire Inc.
©What's Up Hollywood


Mole Valley District Council is asking people to have their say on the Transform Leatherhead project.
A consultation event is held twice a year and involves a Q & A session for residents to voice their concerns.
Key projects to be discussed this time include the renovation of The Swan Centre and the redevelopment of Claire and James house.
The event will be at the Leatherhead Theatre on the 17th of October at 6.30pm.


text:
Yemen: "One of the worst conflicts MSF has seen"
View all News & Stories
By Dr Tammam Aloudat, Médecins Sans Frontières/Doctors Without Borders (MSF) Deputy Medical Director
As we entered the nutrition department of the Mother and Child hospital, a woman sitting at the edge of a bed glanced over at us with suspicion. Strangers usually do not accompany medical staff at this time of the day.
The baby was breathing fast and he seemed in pain. I asked his mother's permission to examine him. She seemed more relaxed once I spoke to her in Arabic.
I told her I worked with MSF, and that we were visiting Ibb to assess the health situation and explore ways to support healthcare facilities, which are struggling under a blockade, airstrikes and the war.
Health situation in Ibb
The hospital director briefed my colleagues as I continued my conversation with the mother. She told me she had come from a village two hours from Ibb.
The worried mother said that her five-month-old was suffering from severe diarrhoea and vomiting. As I examined the baby, the paediatrician told me that the baby was suffering from dehydration but had already improved after one day of treatment.
The mother smiled as she heard the good news, but soon her face turned gloomy again.
When I asked her why, she said she and her husband had had to pay 15,000 YR (around 220 AED) to reach the hospital, and they would have to pay the same to go back; an amount that very few Yemenis can afford and will leave the family in debt for a long time to come.
We have been running an Emergency Surgical Hospital in #Aden, #Yemen, since the beginning of 2015. "The city is still dangerous because of sniper fire and stray bullets. Everyone is armed," says Thierry Goffeau, who has just returned from Yemen after spending three months as an #MSF project coordinator in Aden. "One person shoots because he's happy or for his own amusement and another shoots because he's not happy. As a result, we are still treating patients with #gunshot wounds. You can gather up a handful of stray bullets every day in the #hospital compound where we work in the Sheikh Othman district. Last week, a bullet whizzed by the window of my office. A few days earlier, one came through the wall of the bedroom of one of our surgeons." © Guillaume Binet/MYOP
A photo posted by Médecins Sans Frontières (MSF) (@doctorswithoutborders) on
Aug 10, 2015 at 2:18pm PDT
Desperation in Yemen
This is my second visit to Yemen. Since I last came in 2011 some things have not changed at all, such as the kindness and hospitality of people, but also the long power cuts.
However, many things have changed for the worse. Today, long queues of cars wait in front of petrol stations, and checkpoints have increased. Yemen's quiet nights have turned noisy, filled with the sounds of airstrikes and anti-aircraft guns.
For me, the biggest difference was that the general sense of optimism had turned into desperation and fear for the future. It is sadly a justified fear, as Yemenis are today living through one of the worst armed conflicts MSF has ever seen.
Inhumane conditions
Later, during a trip to one of Ibb's schools where people have sought refuge, we met families who had come to the city fleeing areas witnessing fighting or severe bombardment.
Many had come from Ta'iz and Al Dhale', while others had made the long trip from Sana'a or even Sa'ada.
Around 20 men and several curious boys gathered around us to chat. Most of the boys stood next to their fathers or older brothers, but one five-year-old boy stood next to me, his tiny hand clinging to my shirt.
I was unable to fully concentrate on the discussion as I was reflecting on the inhumane conditions under which Yemen's children are forced to live today.
The family of Fakhira Hameed carry her body to a local cemetery in Ta'iz, after her house was hit by shelling on 24 July.
Most of us in the West can receive mental support after a traumatic event, but the children of Yemen are witnessing a ruthless war, have been forced from their homes and are being deprived of basic needs, healthcare, school and even food, all while their families struggle to survive.
I passed my fingers through the boy's hair in an attempt to offer him some warmth and compassion; two things that are currently in short supply for Yemeni children.
It was the voice of my colleague that brought me back to the conversation. Pointing at me, he was telling a man, "Speak to the doctor."
A tall man with a tired smile, wearing an old shirt and a traditional Yemeni Futah, (a wraparound skirt or kilt) approached me.
"A ruthless war"
He explained that he had a heart condition and described how his health had deteriorated since arriving in Ibb. He smiled when I asked him why it had taken him so long to come to hospital.
He said that even if the consultation was free, he had no money to buy medicine.
Soon, the discussion turned to the topic of food shortages. During Ramadan, the people living in schools had been offered some food for the Iftar (the evening meal that breaks the daily fast) by their neighbours.
Those donations had mostly stopped after Ramadan. International aid organisations are not providing any food aid to the people living in schools, and they cannot afford it by themselves.
Malak Shaher/MSF
MSF international president Dr Joanne Liu visits internally displaced people in Khamer.
Yemeni children, who have already suffered from decades of malnutrition, will suffer more if the world does not provide food and medicine to them.
Yet efforts in this regard are being hindered by the blockade, fighting and constant bombings.
Yemen is experiencing a ruthless war. I hope next time I visit the country, the war will be over.
Until then, MSF will continue to serve the Yemeni people and to project their voices to the world, so that people everywhere else will know the reality beyond the headlines, which talks only about victories, retreats and negotiations.
Find out more about MSF's work in Yemen
Related News & Stories
Syria: Latest attacks on east Aleppo hospitals leave medical care in tatters
Aleppo: "Life has become almost impossible"
Yemen: MSF withdraws its teams from six hospitals in north Yemen
Get a deeper insight into the work of MSF by contacting us.


KMC Interiors provided all window treatments for a prestigious show house. I was amazed and delighted by the service provided. Karen was involved from the design phase, assisting with fabric selection, providing advice and samples, and sharing my enthusiasm and determination to make sure it was just right. Nothing was too much trouble for her.
If you need a friendly, professional and above all extremely knowledgeable person to undertake anything from cushions to antique upholstered furniture then I can't recommend Karen enough. I approached her early last year and have continued to come back ever since!


text:
from __future__ import (absolute_import, division, print_function)
__metaclass__ = type

import os
import json
import pytest
import sys

if sys.version_info < (2, 7):
    pytestmark = pytest.mark.skip("F5 Ansible modules require Python >= 2.7")

from ansible.module_utils.basic import AnsibleModule

try:
    from library.modules.bigip_device_syslog import ApiParameters
    from library.modules.bigip_device_syslog import ModuleParameters
    from library.modules.bigip_device_syslog import ModuleManager
    from library.modules.bigip_device_syslog import ArgumentSpec

    # In Ansible 2.8, Ansible changed import paths.
    from test.units.compat import unittest
    from test.units.compat.mock import Mock

    from test.units.modules.utils import set_module_args
except ImportError:
    from ansible.modules.network.f5.bigip_device_syslog import ApiParameters
    from ansible.modules.network.f5.bigip_device_syslog import ModuleParameters
    from ansible.modules.network.f5.bigip_device_syslog import ModuleManager
    from ansible.modules.network.f5.bigip_device_syslog import ArgumentSpec

    # Ansible 2.8 imports
    from units.compat import unittest
    from units.compat.mock import Mock

    from units.modules.utils import set_module_args

fixture_path = os.path.join(os.path.dirname(__file__), 'fixtures')
fixture_data = {}

def load_fixture(name):
    path = os.path.join(fixture_path, name)

    if path in fixture_data:
        return fixture_data[path]

    with open(path) as f:
        data = f.read()

    try:
        data = json.loads(data)
    except Exception:
        pass

    fixture_data[path] = data
    return data

class TestParameters(unittest.TestCase):
    def test_module_parameters(self):
        args = dict(
            kern_from='err',
            kern_to='info',
        )
        p = ModuleParameters(params=args)
        assert p.kern_from == 'err'
        assert p.kern_to == 'info'

    def test_api_parameters(self):
        p = ApiParameters(params=load_fixture('load_sys_syslog_1.json'))
        assert p.kern_from == 'debug'
        assert p.kern_to == 'emerg'

class TestUntypedManager(unittest.TestCase):

    def setUp(self):
        self.spec = ArgumentSpec()

    def test_update(self, *args):
        set_module_args(dict(
            kern_from='emerg',
            kern_to='debug',
            provider=dict(
                server='localhost',
                password='password',
                user='admin'
            )
        ))

        module = AnsibleModule(
            argument_spec=self.spec.argument_spec,
            supports_check_mode=self.spec.supports_check_mode
        )

        current = ApiParameters(params=load_fixture('load_sys_syslog_1.json'))

        # Override methods to force specific logic in the module to happen
        mm = ModuleManager(module=module)
        mm.update_on_device = Mock(return_value=True)
        mm.read_current_from_device = Mock(return_value=current)

        results = mm.exec_module()

        assert results['changed'] is True

redpajama_set_name: RedPajamaGithub


text:
Sanders: I'm not a single-issue candidate
By Bradford Richardson - 02/21/16 11:23 AM EST
Democratic presidential hopeful Bernie Sanders Bernie SandersBiden to seek minimum wage in COVID-19 proposal Former Sanders spokesperson: Progressives 'shouldn't lose sight' of struggling Americans during pandemic 'I saw my life flash before my eyes': An oral history of the Capitol attack MORE on Sunday dismissed an attack from primary rival Hillary Clinton Hillary Diane Rodham ClintonCan Biden encompass the opposition he embodied? Disney silent on Trump status in Hall of Presidents at Magic Kingdom Biden has an opportunity to win over conservative Christians MORE that he's a "single-issue candidate."
"I haven't the vaguest idea of what she is talking about," Sanders said on CNN's "State of the Union." Sanders then rattled off a list of issues for which he has advocated solutions.
"If she thinks that income and wealth inequality and the fact that the rich get richer while everybody else gets poorer is the only issue, it's not," he said. "We're the only major country on Earth that doesn't have a national healthcare program guaranteeing healthcare to all people – talk about that."
"We need to have free tuition at public colleges and universities, so that our kids today can do well in a global economy," he added. "We need to have the wealthiest people and the largest corporations pay their fair share of taxes. We're working very hard to transform our energy system so we effectively combat climate change."
"We are talking about dozens of issues. I'm not quite sure where Secretary Clinton is coming from."
Tags Bernie Sanders Hillary Clinton Sunday shows


text:
OCCAR.Questions@occar.int
Month reset filter
Programme reset filter
FREMM
FSAF-PAAMS
MALE RPAS
MMCM
LSS PD expansion in Saint-Nazaire
In the wake of LSS PD expansion with the opening of Paris site on 15th February 2019, the new OCCAR satellite of Saint-Nazaire was activated on 3rd June 2019, with the arrival of a first Platform Support Officer, to that will be supplemented in July by a second Staff Member.
Logistic Support Ship Launch
A key milestone in the Logistic Support Ship (LSS) Programme was successfully achieved on 22 June 2018, with the launch of the new 27,200 full load displacement ship at Fincantieri's Riva Trigoso shipyard.
Keel Laying Ceremonies in Riva Trigoso for LSS
A further important target in the Logistic Support Ship (LSS) Programme has been successfully achieved with the keel laying of the aft part of the ship. The respective ceremony took place at FINCANTIERI shipyard in Riva Trigoso (Genova) on 12th July.
OCCAR-EA
53011 Bonn / Germany
+49 (0) 228 5502 0
ISO: 9001:2015
www.tuv.com
Copyright 2020 OCCAR-EA, all rights reserved


Last Thursday, Alvin took Anika and me to Tagaytay because it was a holiday.
The first stop that we went to was the Palace in the Sky.
The last time I was there was in 2006, I think. My ex-boyfriend took me there and I was kinda weirded out bakit ang daming tao eh dilapidated na yung place.
Since it was like 8 years ago, I thought Palace in the Sky improved. Hindi naman. I'll show you later.
The view was fantastic though given that we were on the top most part of Tagaytay.
When you reach the entrance of the Palace in the Sky, you are to pay an entrance fee. Sana they ask na lang for a bigger amount and use it to rehabilitate the place.
Upon entering, your choice is to walk up the hill to the top or ride a jeepney. Ayaw kong malusaw ang make-up ko so we rode a jeepney to the top for Php 40 for the three of us. That was also the first time that Anika experienced riding a jeep. Di niya masyadong type but when we were going down na, she enjoyed it already.
They really ought to fix the place especially the railings. Pwedeng pwede mahulog ang kahit sino diyan. Horror.
There were stalls there that were selling Tagaytay souvenirs. I got Anika an umbrella.
Alvin wanted to go check out the "palace". Ako hindi na because I was scared. By the looks of the structure, I felt it wasn't safe to go in anymore.
Imelda Marcos decided to build a mansion on top of Mount Gonzales, which was at that time a land owned by the government. The summit before was used by the Bureau of Air Transport as a radar station. Construction of the Palace in the Sky began in 1981. The construction of the mansion was a tedious one as roads were built for supplies to reach the top of the 2,500 ft Mount Gonzales. The mountain was leveled by bulldozers and farmers living in the mountainside were asked to relocate. Construction of the mansion was hastened when US President Ronald Reagan announced his intention to visit the Philippines in November 1983. It was reported that the Marcos administration planned to accommodate Reagan at the Palace in the Sky. Post-People Power Revolution critics of the Marcos administration described the unfinished mansion as a symbol of the Marcos administration's excess.
Grabe. They didn't finish pala building the mansion.
Sayang right? I wish the government will fix it up. Sayang the fantastic view talaga. I hope they can put restaurants there or a nice park where people can have picnics or just simply hang out.
By the parking area, you'll see a terrible canteen.
Literally, it's Terrible's Canteen talaga.
« Mommy Fleur Discovers Virgin Beach Resort.


1 Kings 8:39 39 then hear in heaven Your dwelling place, and forgive, and act, and give to everyone according to all his ways, whose heart You know (for You alone know the hearts of all the sons of men).
God knows the hearts of all people and He gives to men according to his deeds. Pray that God's hand will be free in America to help law enforcement apprehend law breakers and that there be justice in the courts according to God's understanding of men's hearts.


text:
Q: Background Image incorreto em IPhone Estou usando:
    <section class="parallax calltoaction section_padding_100"
    style="
        background-image: url('images/models_cover/landscapes.jpg');
        background-size:100% 100%;
        background-repeat: no-repeat;
    ">

Ao acessar pelo chrome, firefox e safari via desktop funciona perfeitamente como na imagem.

Ao acessar via navegador Android Chrome, funciona perfeitamente também.

Mas, ao acessar via iPhone com o navegador padrão ou Chrome, fica da seguinte forma.

Já troquei o background-size:  por cover e container, mas, fica na mesma.

A: Pelo que dá para ver no seu HTML, você está usando uma classe chamada parallax. Provavelmente essa classe está usando o atributo background-attachment: fixed, pois é através dessa propriedade que se faz o efeito mais simples de parallax.
O que acontece é que esse tipo de efeito não é performático em dispositivos móveis, normalmente smartphones, e a maioria dos navegadores desabilitam-o por padrão. Porém, em dispositivos com iOS ocorre o erro descrito por você, que ao invés de desabilitar a propriedade background-attachment com o valor fixed, ele faz uma espécie de zoom que não é o esperado por nós.
A solução que pode ser feita no caso de uma página responsiva, onde se deseja obter o efeito de parallax no desktop e desabilitar no mobile, é através de JavaScript, modificar o valor do atributo background-attachment: fixed para scroll em todos os elementos que utilizem a classe .parallax. Ou então, se você se baseia em resolução para saber se é mobile ou desktop, você pode definir isso em um  media query no CSS.

A: Tenta com o seguinte codigo, funcionou perfeitamente na minha aplicação

html {
    background: url(http://placebear.com/1200/800) no-repeat center center fixed;
    background-size: cover;
    height: 100%;
    overflow: hidden;
}
body{
    height:100%;
    overflow: scroll;
    -webkit-overflow-scrolling: touch;
}

redpajama_set_name: RedPajamaStackExchange


text:
Q: How to convert a String with coordinates to a List in dart I want to convert a string with value "[0, 0]" to a list with value [0, 0]. Somewhat like casting a string to a list.
Code:
void main(){
  String cords_string = "[0, 0]";
  List<int> cords_list = ;//i want to make this have a value of [0, 0] or whatever value cords_string has
}

A: You can use json.decode from dart:convert to get a List<dynamic>, and from there you can convert it to a List<int>. This is one way to do it:
import 'dart:convert';

void main() {
  String cords_string = "[0, 0]";
  List<int> cords_list = [...json.decode(cords_string)];
  print(cords_list);
}

Alternatively you could use List<int>.from(json.decode(cords_string)) instead.


text:
Kuark Forest
Citizen Science Survey Camp – Spring 2018
The Goongerah Environment Centre (GECO), based in far east Gippsland is hosting a citizen science survey over the Melbourne Cup long weekend in November (November 3 – 6, 2018).
"You will learn from the dedicated and passionate ecologists and activists at GECO whose citizen science campaign is saving forests from logging. We'll be based in and around Goongerah, including surrounding high conservation value and old growth forests, and the iconic Kuark forest".
Continue reading "Citizen Science Survey Camp – Spring 2018" →
Emerald Link film to be launched
East Gippsland is the only place on mainland Australia with continuous forests from the alps to the sea.
The vision to see the Emerald Link created seeks to protect this precious landscape and biodiversity and create a viable economic future based on nature tourism.
Last year, Goongerah Environment Centre (GECO) shot a stunning film about the proposal. Produced by award winning cinematographer David Franjic from Colour Chorus. The Emerald Link film captures the wild beauty of East Gippsland's forests and the stories of the people who love it. It is being launched on July 7.
Continue reading "Emerald Link film to be launched" →
Protection for some of Kuark forest a welcome first step
Following a long community campaign, the Andrews government that it will increase protection of the Kuark forest in East Gippsland.
As part of the extension of the Victorian Regional Forests Agreements (RFAs), the government has announced protection for the Kuark forest. The Kuark is a classic ecotone forest that exists on the interface between the cooler forest types of Gippsland and the sub tropical forests that stretch up the eastern seaboard. There are plans to create a 'coast to mountains' walking trail through this forest.
Spokesperson for the Goongerah Environment Centre Office (GECO), Ed Hill, said "we welcome the government's announcement that parts of the Kuark forest will now be protected. This is a good step in the right direction and we look forward to working with the government to ensure the Kuark forest is formally incorporated into the Errinundra National Park along with other forest areas of high environmental value, within this term of government.
"Today's announcement does not provide full protection for the iconic Kuark forest, some areas have not been included, we will continue to advocate and campaign for their protection."
The full reaction from GECO can be found here and Friends of the Earth here. The minister's announcement is here.
A new long distance walking trail for East Gippsland
The campaign for the Emerald Link park in East Gippsland aims to protect the more-or-less intact ecosystems that run from the coast to the mountains. A long distance walking trail is an integral part of the proposal. The proposed Sea to Summit Forest Trail would create a network of walking tracks linking the coastal town of Bemm River and the existing Wilderness Coast walk to the summit of Mount Ellery, the highest mountain in far East Gippsland.
Continue reading "A new long distance walking trail for East Gippsland" →
Kuark forest protected through Court injunction
The Kuark forest is located in far East Gippsland, Victoria. This magnificent forest is home to rare rainforest and endangered animals. Sections of it are currently on logging schedules and could be cut at any moment. An access track has been cut into the first coupe.
In response, campaigners have set up a camp in the forest to oppose logging. In response, the Victorian government announced it would increase the protection given to old growth forests, but activists are not prepared for any of the coupe to be logged.
On Wednesday November 1st, a Supreme Court injunction has prevented logging from starting in the forest. Campaigners are celebrating this temporary protection.
Further information is available on the GECO website.


This takes 2 days to make, so leave yourself enough time to soak the fruit the night before (see below) and then at least 3hrs for baking the next day or longer if you choose to make more than one cake from this mix. This can reap you 1 large cake or 2/3 smaller ones.
For a truly Zobird's twist it only felt right to use Sailor Jerry or added spice…and booze-impact!
Chop up the glacé cherries…I used a full pot…Mmmmm!
Add these to all the dried fruits and mix it up – Even distribution is the key!
Empty everything in to a ceramic bowl and leave to cool.
Add the almonds to the dry and start alternating between the pre-soaked/blitzed fruit and the dry on a slow speed.
After 2hours of the cake cooling gradually in the foil and tea towel, remove from its toasty parcel…It may still be warm! Plus it'll smell LUSH!!!
Prepare 2 large sheets of foil and a new piece of greaseproof…place the cake in the centre and wrap it up like a tasty parcel…I went for the 'bring it all to the middle' approach as you will need to access this over the next 8weeks or so.
At this point the cakes can be stored in a cool place in the containers until you decide to 'feed' the cake. i will be feeding mine once a week for the next 7 weeks before icing them…and marzipan for my mum!
To feed the cake you can brush the top and let it sink in but using a cocktail stick to stab holes in to the cakes will help the booze soak further through. I'm going to be putting about an extra tablespoon per week in to the cake.
There's still time to make this and feed it plenty before Christmas, so why not give it a go.


Today in Heritage History, March 31, 1870, Texas is the last Confederate state to be readmitted to the Union. It took Texas about 12 attempts, over a six-year period to (finally) pass the G.E.D., which was the Union's criteria for re-admission.
Today in Heritage History, March 30, 1867, the U.S. buys Alaska from Russia for $7.2 million. Russia was originally asking $10.5 million for the territory, but agreed to the discounted price if government officials in the U.S. promised to keep the Palin's and never tell them where Russia "moved to".