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LMO3 promotes hepatocellular carcinoma invasion, metastasis and anoikis inhibition by directly interacting with LATS1 and suppressing Hippo signaling
Yang Cheng1,
Tianlu Hou1,
Jian Ping1,
Tianyang Chen1 &
Baobing Yin2
In this research, we aimed to investigate the biological functions of LIM domain only 3 (LMO3) in hepatocellular carcinoma (HCC) and uncover the underlying molecular mechanism in it.
HCC tissue microarray (n = 180) was used to analyze the correlation between LMO3 expression and clinicopathological findings. In vitro transwell matrigel invasion assay and annexin V anoikis assay in HCC cells were conducted to investigate LMO3 related biological functions. In vivo intrahepatic and lung metastasis models were used to determine the role of LMO3 in HCC metastasis. Quantitative real-time PCR, western blotting and immunohistochemical staining were performed to investigate the expression and mechanism of LMO3 in HCC.
We found that the expression of LMO3 was significantly upregulated in HCC tissues, and it was closely related to clinicopathological findings and patient prognoses. Knockdown of LMO3 suppressed the invasion and anoikis inhibition of HCC cells in vitro. Meanwhile, the metastasis of SMMC-7721 cells was also suppressed by LMO3 knockdown in vivo. Furthermore, we found that LMO3 knockdown increased the phosphorylation of YAP and LATS1, and decrease Rho GTPases activities. LMO3 directly interacted with LATS1, and thus suppressed Hippo signaling. Recombinant LMO3 (rLMO3) protein administration decreased the phosphorylation of YAP and LATS1, and increased Rho GTPases activities. The inhibitors of the Hippo pathway abrogated rLMO3 protein-induced HCC cell invasion and anoikis inhibition.
These results suggest that LMO3 promotes HCC cell invasion and anoikis inhibition by interacting with LATS1 and suppressing Hippo signaling. LMO3 may serve as a potential therapeutic target for HCC in future.
Hepatocellular carcinoma (HCC) is one of the most common malignancies and a leading cause of cancer-related mortality worldwide [1, 2]. The risk factors for HCC, such as hepatitis B virus or hepatitis C virus infection, are well documented. Tumor metastasis are the main cause of death in patients with HCC. Recent studies suggest that tumor metastasis is a complex process affected by multiple procedures and multiple mediators in human cancers [3, 4]. However, the underlying molecular mechanism in HCC metastasis still remains poorly understood. Tumor metastasis may be affected by many intracellular signaling molecules and extracellular components, such as cytokines, neurotransmitters or the extracellular matrix [5, 6].
The Hippo pathway is an evolutionarily conserved signaling module that plays critical roles in liver size control and tumorigenesis [7, 8]. In mammals, the Hippo pathway is a kinase cascade, wherein Macrophage Stimulating 1/2 (MST1/2), in a complex with its regulatory protein Salvador Family WW Domain Containing Protein 1 (SAV1), phosphorylates and activates Large Tumor Suppressor Kinase 1/2 (LATS1/2). Yes-Associated Protein (YAP)/ Tafazzin (TAZ) can be phosphorylated and inactivated by active LATS1/2; when it is restrained in the cytoplasm, it loses its transcriptional activation of pro-proliferation and anti-apoptosis genes. The Hippo pathway can be activated by many biological factors including contact inhibition, mechanical strain on the cell, cell polarity/adhesion molecules, other signaling pathways and cellular metabolic status [9]. Cytokinesis failure has recently been shown to trigger the Hippo pathway [10]. Nevertheless, the link between LIM domain only 3 (LMO3) and the Hippo pathway has not been reported.
The LMO protein family includes four members: LMO1, LMO2, LMO3 and LMO4. Although LMO proteins lack DNA binding activity, some reports indicate that they are involved in the transcriptional regulation of target genes in collaboration with other transcription factors. Genetic analyses indicate that LMO1 and LMO2 contribute to the genesis of immature and aggressive T-cell leukemia [11]. LMO4 was implicated in the development of breast cancer [12, 13]. LMO3 was reported to form a complex with neuronal-specific basic helix-loop-helix (bHLH) transcription factor Helix-Loop-Helix protein 2 (HEN2), which was also expressed at higher levels in unfavorable neuroblastoma than in the favorable type. Moreover, LMO3 has been reported to play important roles in some types of cancer, including neuroblastoma [14, 15] and lung cancer [16, 17].
In this research, we found that the expression of LMO3 was significantly upregulated in HCC tissues. LMO3 expression was closely related to clinicopathological findings or patient prognoses. Knockdown of LMO3 suppresses the invasion, metastasis and anoikis inhibition of HCC cells. Further, the effects of LMO3 on the biological behaviors of HCC cells are dependent on the suppression of Hippo signaling.
Human HCC cell lines, including HCCLM3, HepG2, Huh-7, MHCC-97H, MHCC-97 L, SK-Hep1, SMMC-7721, SNU-423 and SNU-449 were purchased from Cell Bank of the Chinese Academy of Sciences. Dulbecco's modified Eagle's medium (DMEM) contained 10% (v/v) fetal calf serum (FCS) and 1% antibiotics was used. Cells were incubated at 37 °C in a humidified incubator under 5% CO2 condition.
Clinical samples
Clinical human HCC (16 cases) and corresponding non-cancerous liver (CNL) tissues (12 cases), in which 12 cases were paired, were obtained from Shanghai University of Traditional Chinese Medicine Affiliated Shuguang Hospital. Additionally, Human tissue microarray contained 180 cases of HCC samples was bought from Alenabio.
Ethics, consent and permissions
All human samples were obtained with informed consent. The protocols were approved by the ethical review committee of the World Health Organization Collaborating Center for Research in Human Production (authorized by the Shanghai Municipal Government).
Quantitative real-time PCR
Total RNA was extracted by Trizol (Takara), and reversely transcribed by PrimeScript RT-PCR kit (Perfect Real Time). Quantitative real-time PCR analyses were performed by SYBR Premix Ex Taq (Takara) on a 7500 real-time PCR system (Applied Biosystems), with recommended thermal cycling settings: one initial cycle at 95 °C for 30 s followed by 40 cycles of 5 s at 95 °C and 31 s at 60 °C. Primer sequences used for human LMO3, CTGF, ANKRD1 and CYR61 detection were shown in Additional file 1: Table S1.
All tissue samples were fixed in phosphate-buffered neutral formalin, embedded in paraffin, and cut into 5 μm thick sections. The sections were deparaffinized and rehydrated, incubated with 0.3% hydrogen peroxide/phosphate-buffered saline for 30 min, and blocked with 10% BSA (Sangon). The antibody for LMO3 (Abcam) was used to incubate the slides at 4 °C overnight with optimal dilution. HRP (rabbit) second antibody (Huabio) was used to incubate above slides at room temperature for 1 h. The slides were labeled with DAB substrate liquid (Thermo Scientific) and counterstained by hematoxylin. All the sections were photographed with a microscope (Carl Zeiss). Scoring was designated according to the ratio and intensity of positive-staining cells: 0–5% scored 0; 6–40% scored 1; 41–70% scored 2; more than 70% scored 3. The final score was defined as low or high expression group as follows: score 0–1, low expression, score 2–3, high expression. All scores were determined independently by more than two senior pathologists in a blinded manner.
Western blotting and GTPase pull-down assays
Cells were lysed in lysis buffer. Then the proteins were separated by SDS-PAGE under reducing condition. The membranes were blocked in phosphate-buffered saline/Tween-20 containing 5% BSA, then incubated by the antibodies for LMO3 (Abcam), phospho-YAP (Cell Signaling), total-YAP (Cell Signaling), phospho-LATS1 (Cell Signaling), total-LATS1 (Cell Signaling), GAPDH (Huabio) and species-specific secondary antibodies separately. The membranes were detected by Odyssey imaging system (LI-COR). GTPase pull-down assays were performed according to standard procedures as described [18].
siRNA or shRNA transfection
Small interfering RNAs duplexes for LMO3 used in this study was produced by Genepharma. Transfection steps were performed according to the manufacture's protocols. The sequences of siRNA were designed as: si-LMO3–1: F: GGACUACGAGGAAGGUUUAdTdT, R: UAAACCUUCCUCGUAGUCCdTdT; si-LMO3–2: F: GCUGCAACCGAAAGAUCAAdTdT, R: UUGAUCUUUCGGUUGCAGCdTdT. Further, shRNA sequence was designed as: sh-LMO3: F: GATCCGTACACTAAAGCTAATCTT ATCTTCCTGTCAGAATAAGATTAGCTTTAGTGTACTTTTTG, R: AATTCAAAAAGTACACTAAAGCTAATCTTATTCTGACAGGAAGATAAGATTAGCTTTAGTGTACG. The structure of pGreenPuro used for shRNA and vector construction was shown in Additional file 2: Figure S2.
rLMO3 protein and inhibitors
Recombinant LMO3 (rLMO3) protein was purchased from Abnova. The inhibitor of Hippo (Verteporfin and Peptide 17) were purchased from Selleck.
In vitro invasion assay
MHCC-97H or SMMC-7721 cells were detached by 0.25% trypsin/0.01% EDTA in 1 × PBS and resuspended in serum-free DMEM medium. 2 × 104 cells in 100 μl were added into matrigel (BD)-coated inserts (Millipore) seated on the 24-well plate. Then DMEM medium contained 5% FBS was added into the bottom chamber. After the cells were incubated at 37 °C for 48 h, filters were fixed and stained with 0.1% (w/v) Crystal Violet. Non-invading cells were removed firstly, and invading cells were counted under a microscope at a magnification of 400×. About 3 grids per field were counted. All of the experiments were repeated twice.
Anoikis assays
5 × 105 MHCC-97H or SMMC-7721 cells were cultured on poly-HEMA treated 12-well plates at 37 °C for 48 h. Then the adherent cells were detached and harvested in complete DMEM medium and centrifuged at 1000 rpm/5 min. The cells were washed with 1 × PBS and incubated with 100 μl binding buffer containing 3.5 μl Annexin V and 3.5 μl propidium iodide (PI) at room temperature for 15 min. All of the cells were analyzed by flow cytometry (BD).
Edu assay
1 × 106 MHCC-97H or SMMC-7721 cells were seeded into 6-well plates. 50 μM of Edu from Edu Apollo® 488 In Vitro Flow Cytometry Kit (RiBoBio) was added into the plates 2 h before harvesting the cells. Cells were collected and centrifuged at 1000 rpm/5 min, and supernatant was removed. For fixation, 4% paraformaldehyde was added into the cells and incubated for 15 min, and washed once by 1 × PBS. Then cells were resuspended in Tris buffer saline with 0.5% Triton X-100 and incubated for 10 min, and washed again with 1 × PBS. Amounts of 500 μl staining solution with Apollo® 488 fluorescent azide was added into cells, incubated for 10 min, and then rinsed twice with Tris buffer saline with 0.5% Triton X-100. All of the cells were analyzed by flow cytometry (BD).
In vivo metastasis assays
2 × 106 SMMC-7721 cells infected with sh-LMO3 or control, were detached and suspended in 30 μl serum-free DMEM/matrigel (1:1) for each BALB/c-nu/nu mouse. Through a 1 cm transverse incision in the upper abdomen under anesthesia, each mouse (6 weeks, male, 10 in each group) was orthotopically inoculated in the left hepatic lobe with a microsyringe. Meanwhile, 1 × 106 SMMC-7721 cells infected with sh-LMO3 or control were injected intravenously into nude mice (6 in each group). Mice were sacrificed after 6 weeks. The livers and lungs were dissected, fixed with phosphate-buffered neutral formalin and prepared for standard histological examination. All of mice were manipulated and housed according to protocols approved by the Shanghai University of Traditional Chinese Medicine Animal Care Commission. All animals received humane care according to the criteria outlined in the "Guide for the Care and Use of Laboratory Animals" prepared by the National Academy of Sciences and published by the National Institutes of Health.
Co-immunoprecipitation
For intracellular immunoprecipitation, Huh-7 cell lysates transfected with HA-tagged LMO3 or vector control were subjected to immunoprecipitation with anti-HA monoclonal antibody (Millipore) or control IgG. Then the immunoblotting with anti-LATS1 or YAP antibodies was performed.
Values are expressed as the mean ± standard error of the mean. Statistical analyses were performed using SPSS 16.0 for windows. Survival time was analyzed with the Kaplan-Meier method. The association between LMO3 expression and the clinicopathological features of HCC patients was evaluated using Pearson's Chi-square test. One-way analysis of variance was used for comparison between groups. P < 0.05 was considered to indicate a statistically significant difference.
LMO3 is upregulated in HCC, and closely related with the clinicopathological findings and patient prognoses
To investigate the expression level of LMO3 in HCC tissues, we collected 16 cases HCC and 12 cases CNL tissues. Through Quantitative real-time PCR, we found that the expression of LMO3 was significantly upregulated in HCC tissues (Fig. 1a). In 12 paired HCC and CNL tissues, the mRNA expression of LMO3 was found to be significantly upregulated in HCC tissues (Fig. 1b). Furthermore, in 5 paired HCC and CNL tissues, the protein expression of LMO3 was also found to be significantly upregulated in HCC tissues (Fig. 1c).
LMO3 expression is upregulated in HCC and closely related with clinicopathological findings or patient prognoses. a The mRNA expression level of LMO3 in 16 cases HCC and 12 cases corresponding noncancerous liver (CNL) tissues. b The mRNA expression level of LMO3 in 12 paired HCC and CNL tissues. c The protein expression level of LMO3 in 5 paired HCC and CNL tissues. **P < 0.01. d The immunohistochemical staining of LMO3 in HCC and CNL tissues. Scale bars: 10 μm. e By analyzing HCC tissue microarray (n = 180), the expression of LMO3 was found to be upregulated in 75.40% HCC tissues. f The immunohistochemical staining of LMO3 in HCC, thrombus, CNL and normal liver tissues. Scale bars: 5 μm. g Western blotting analysis of LMO3 expression in HCC, thrombus, CNL and normal liver tissues. GAPDH was used as a loading control. h and i Kaplan-Meier analysis of overall survival (OS) (f, P = 0.019) and disease-free survival (DFS) (g, P = 0.179) for the expression of LMO3
Then we used an HCC tissue microarray (n = 180) to detect the correlation between LMO3 expression and clinicopathological findings and patient prognoses (Additional file 3: Table S2). We found that the expression of LMO3 was upregulated in 75.40% HCC tissues (Fig. 1e). The upregulated LMO3 expression in HCC tissues was further confirmed by immunohistochemical staining (Fig. 1d). Also, we found that the expression of LMO3 was closely related to patient tumor encapsulation, thrombus, vascular invasion and TNM stage (Table 1). To investigate the relevance of LMO3 with HCC metastasis, we collected HCC, tumor thrombus, CNL and normal liver tissues from the same patients and detected LMO3 expression in these tissues. Through immunohistochemical staining and western blotting, we found that LMO3 expression was higher in the HCC and tumor thrombus tissues, and lower in the CNL and normal liver tissues (Fig. 1f, g). Additionally, we found that the high expression of LMO3 was positively correlated with poor overall patient survival (OS) (P = 0.019), while it was not correlated with patient disease-free survival (DFS) (P = 0.179) (Fig. 1h, i).
Table 1 Correlation of the clinicopathological findings with LMO3 expression
Knockdown of LMO3 suppresses the invasion and anoikis inhibition of HCC cells in vitro
To investigate the biological functions of LMO3 in HCC, we detected the expression level of LMO3 in 9 HCC cell lines. As shown in Fig. 2a, we found that LMO3 had high expression levels in MHCC-97H and SMMC-7721 cells. Then we selected MHCC-97H and SMMC-7721 cells and knocked down LMO3 by using siRNA (labeled as si-LMO3–1 and si-LMO3–2). Through western blotting analyses, we found that LMO3 was successfully silenced in MHCC-97H (Fig. 2b) and SMMC-7721 cells (Fig. 2c).
LMO3 knockdown suppresses the invasion and anoikis inhibition of HCC cells in vitro. a Expression of LMO3 in HCC cell lines, including HCCLM3, HepG2, Huh-7, MHCC-97H, MHCC-97 L, SK-Hep1, SMMC-7721, SNU-423 and SNU-449 cells. b and c The protein expression level of LMO3 in MHCC-97H (b) and SMMC-7721 (c) cells, which were infected with siRNA of LMO3 or NC. d Representative photos of invaded MHCC-97H cells infected with siRNA of LMO3 or NC. Statistical analysis of invaded MHCC-97H cells in the three groups is shown below. Scale bars: 10 μm. e Representative photos of invaded SMMC-7721 cells infected with siRNA of LMO3 or NC. Statistical analysis of invaded SMMC-7721 cells in the three groups is shown below. Scale bars: 10 μm. f Flow cytometry analysis of anoikis of MHCC-97H cells infected with siRNA of LMO3 or NC. Flow cytometry statistical analysis of apoptotic MHCC-97H cells in the two groups is shown below. g Flow cytometry analysis of anoikis of SMMC-7721 cells infected with siRNA of LMO3 or NC. Flow cytometry statistical analysis of apoptotic SMMC-7721 cells in the two groups is shown below. **P < 0.01, NS, no significant
Then we investigated the role of LMO3 in the invasion of HCC cells. By transwell matrigel invasion assay, it was found that knockdown of LMO3 suppressed the invasiveness of MHCC-97H and SMMC-7721 (Fig. 2d, e) cells after 48 h. Furthermore, through annexin V anoikis assay, we found that the knockdown of LMO3 promoted the anoikis of MHCC-97H and SMMC-7721 (Fig. 2f, g) cells after 48 h. Additionally, by Edu assay, we found that the proliferation of MHCC-97H and SMMC-7721 cells was not affected by LMO3 knockdown at 24-, 48- and 72-h time points respectively (Additional file 4: Figure S1a, b).
Knockdown of LMO3 suppresses the intrahepatic and distant metastasis of HCC in vivo
To further investigate the role of LMO3 in cancer progression in vivo, sh-LMO3 and control cells (SMMC-7721) were orthotopically injected into nude mice to assess intrahepatic metastases and intravenously transplanted to assess distant metastases. After 6 weeks, we found that the intrahepatic metastatic nodules in the sh-LMO3 group were obviously less than in the vehicle group (Fig. 3a, b). Mice survival in the LMO3-silenced group was significantly increased compared to the vehicle group (Fig. 3c). Histological staining of the liver tissues indicated that mice transplanted with sh-LMO3 cells had fewer intrahepatic metastatic nodules than those transplanted with control cells (Fig. 3d, e). Meanwhile, the pulmonary metastatic nodules in mice transplanted with sh-LMO3 cells were less than those transplanted with control cells (Fig. 3f, g).
LMO3 knockdown suppresses the metastasis of HCC cells in vivo. a Representative photos of liver tissues from mice orthotopically inoculated with LMO3 silenced SMMC-7721 and control cells (n = 5 each group). b Statistical analysis of intrahepatic metastatic tumor volume is shown right. c Mice survival of LMO3 silenced SMMC-7721 and control group (n = 10 each group) (P < 0.001). d Representative images of H&E staining in liver tissues from mice orthotopically inoculated with LMO3 silenced SMMC-7721 and control cells (n = 5 each group). Scale bars: 10 μm. e Statistical analysis of intrahepatic metastatic focis in the two groups is shown below. f Pulmonary metastases were detected by H&E staining (n = 6 each group). Scale bars: 10 μm. g Statistical analysis of numbers of pulmonary metastatic nodules is shown below. **P < 0.01
LMO3 knockdown increases the phosphorylation of YAP/LATS1 signaling and decreases Rho GTPases activities
We further investigated the underlying mechanism of LMO3 in HCC. Through western blotting analyses, we found that the Hippo signaling pathway played important roles in HCC invasion and metastasis. Interestingly, knockdown of LMO3 in MHCC-97H or SMMC-7721 cells significantly increased the phosphorylation of YAP. Furthermore, the phosphorylation of LATS1, a core component of the Hippo pathway, was also increased by silencing LMO3 (Fig. 4a, b). Then LMO3 knockdown and control MHCC-97H or SMMC-7721 cells were serum starved for 24 h. The activities of RhoA and Cdc42 were measured by pull-down assays. We found that RhoA and Cdc42 activities were suppressed by LMO3 knockdown. Rac1 activity had no obvious change in LMO3 knockdown and control MHCC-97H or SMMC-7721 cells (Fig. 4c, d).
Knockdown of LMO3 increases the phosphorylation of YAP/LATS1 and decreases Rho GTPases activities. a and b Western blotting analysis of phospho-YAP, YAP, phospho-LATS1 and LATS1 in LMO3 knockdown and control MHCC-97H (a) or SMMC-7721 cells (b). Statistical analyses of phospho-YAP/YAP and phospho-LATS1/LATS1 densitometry are shown below. c and d MHCC-97H (c) or SMMC-7721 (d) cells were serum starved for 24 h and the activities of RhoA, Cdc42 and Rac1 were measured by pull-down assays in LMO3 knockdown and control cells. Statistical analyses of active-RhoA/total-RhoA and active-Cdc42/total-Cdc42 densitometry are shown below. e The mRNA levels of CTGF, ANKRD1 and CYR61 in LMO3 knockdown and control MHCC-97H cells. **P < 0.01. f Western blotting analysis of CTGF, ANKRD1 and CYR61 in LMO3 knockdown and control MHCC-97H cells
We also detected the mRNA levels of canonical YAP target genes, including Connective Tissue Growth Factor (CTGF), Ankyrin Repeat Domain 1 (ANKRD1), and Cysteine Rich Angiogenic Inducer 61 (CYR61). It was found that CTGF, ANKRD1 and CYR61 mRNA or protein levels were significantly suppressed in LMO3-silenced MHCC-97H cells (Fig. 4e). These results suggested that the Hippo pathway was activated in LMO3-silenced HCC cells.
LMO3 directly interacts with LATS1, and recombinant LMO3 protein administration decreases the phosphorylation of YAP/LATS1 signaling and increases Rho GTPases activities
Co-immunoprecipitation experiments were performed to investigate whether LMO3 was directly associated with the components of the Hippo pathway. First, Huh-7 was transfected with HA-tagged LMO3 or vector control. We found that immunoprecipitates of LMO3 from Huh-7 cells contained LATS1, but no YAP (Fig. 5a). Furthermore, we added recombinant LMO3 (rLMO3) protein into Huh-7 cells, in which LMO3 had a low expression level. The results showed that the phosphorylation of YAP or LATS1 was significantly suppressed after treatment with rLMO3 protein (Fig. 5b, c). Additionally, RhoA and Cdc42 activities were increased by rLMO3 protein administration, while Rac1 activity had no obvious change (Fig. 5d, e).
LMO3 directly interacts with LATS1, and recombinant LMO3 protein administration suppresses the phosphorylation of YAP/LATS1 and increases Rho GTPases activities. a Co-immunoprecipitation of LMO3 with LATS1 or YAP. Huh-7 cell lysates transfected with HA-tagged LMO3 or vector control were subjected to immunoprecipitation with anti-HA monoclonal antibody or control IgG, followed by immunoblotting with anti-LATS1 or YAP antibodies. The input control on the right panel shows the levels of transfected HA-LMO3 and LATS1 or YAP in HA-tagged LMO3 or vector control transfected cells. b and c Western blotting analysis of phospho-YAP, YAP, phospho-LATS1 and LATS1 in recombinant LMO3 (rLMO3) protein treated and control Huh-7 cells (b). Statistical analyses of phospho-YAP/YAP and phospho-LATS1/LATS1 densitometry are shown right (c). d and e Huh-7 cells were serum starved for 24 h and the activities of RhoA and Cdc42 were measured by pull-down assays in rLMO3 protein treated and control cells (d). Statistical analyses of active-RhoA/total-RhoA and active-Cdc42/total-Cdc42 densitometry are shown right (e). f The mRNA levels of CTGF, ANKRD1 and CYR61 in rLMO3 protein treated and control Huh-7 cells. *P < 0.05, **P < 0.01. g Western blotting analysis of CTGF, ANKRD1 and CYR61 in rLMO3 protein treated and control Huh-7 cells
Meanwhile, we also found that CTGF, ANKRD1 and CYR61 mRNA or protein levels were significantly increased in rLMO3 protein treated Huh-7 cells (Fig. 5f). Thus, we conclude that LMO3 interacts with LATS1, increases Rho GTPases activities and suppresses the Hippo signaling pathway.
Hippo pathway inhibitors could abrogate recombinant LMO3 protein-induced HCC cell invasion and anoikis inhibiton
By using Verteporfin (the inhibitor of YAP-TEAD) and Peptide 17 (YAP-TEAD inhibitor I, the inhibitor of YAP-TEAD), we further investigated the effects of LMO3 on HCC cell invasion and anoikis inhibition. rLMO3 protein was added into Huh-7 and SNU-423 cells, in which LMO3 had low expression levels. Then, Verteporfin and Peptide 17 were added after 2 h. We found that Verteporfin and Peptide 17 abrogated rLMO3 protein-induced Huh-7 or SNU-423 cell invasion (Fig. 6a, b). Meanwhile, the anoikis inhibition of Huh-7 or SNU-423 cells induced by rLMO3 protein was also abrogated by these inhibitors (Fig. 6c, d).
Hippo pathway inhibitors abrogate recombinant LMO3 protein induced HCC cell invasion and anoikis inhibiton. a and b Huh-7 and SNU-423 cells were treated with 50 nM rLMO3 protein, 50 nM rLMO3 protein plus 50 nM Verteporfin (the inhibitor of YAP-TEAD), 50 nM rLMO3 protein plus 50 nM Peptide 17 (the inhibitor of YAP-TEAD) respectively. The invaded Huh-7 (a) and SNU-423 (b) cells were analyzed after 48 h. Scale bars: 10 μm. c and d Huh-7 and SNU-423 cells were treated with 50 nM rLMO3 protein, 50 nM rLMO3 protein plus 50 nM Verteporfin, 50 nM rLMO3 protein plus 50 nM Peptide 17 respectively. The apoptotic Huh-7 (c) and SNU-423 (d) cells were analyzed after 48 h. *P < 0.05, **P < 0.01. e LMO3 promoted HCC cell invasion and anoikis inhibition by interacting with LATS1 and suppressing Hippo signaling
These results indicate that LMO3 promoted HCC cell invasion and anoikis inhibition by interacting with LATS1 and suppressing Hippo signaling (Fig. 6e).
In recent years, studies on LMO3 in some types of cancers have been reported [14,15,16,17]. Nevertheless, the detailed biological functions and related mechanism of LMO3 in HCC were first investigated in this research. We found that LMO3 expression was closely related with tumor metastasis related clinicopathological findings and patient prognoses. Our experiments in vitro and in vivo revealed that knockdown of LMO3 suppresses cell invasion and anoikis inhibition in HCC. All of the above data suggested that LMO3 is involved in the invasion, metastasis and anoikis inhibition of HCC cells.
Invasion and metastasis are major concerns during the progression of cancer [19,20,21,22,23]. The Hippo pathway is known to be pivotal in modulating the invasion and metastasis of tumor cells [24,25,26,27]. YAP is known to contribute to metastasis via multiple mechanisms. YAP interacts with TEAD and FOS in the nucleus, and reprograms gene expression to induce epithelial-mesenchymal transition (EMT). YAP also antagonizes E-cadherin junction assembly by regulating actin cytoskeleton organization and contributes to EMT. Furthermore, YAP activation supports stiffening of the extracellular matrix of cancer-associated fibroblasts (CAFs) to enhance YAP nuclear localization in tumor cells. Such interplay between cancer cells and CAFs might amplify the effects of YAP during tumorigenesis [28,29,30,31].
LMO3 has been proved to promote cell invasion or proliferation through Akt-mTOR/GSK3β signaling in gastric cancer [32]. In this research, we found that knockdown of LMO3 increased the phosphorylation of YAP and LATS1, and thus restrained them in the cytoplasm where they lost their transcriptional activation. Furthermore, these results were confirmed by detecting Rho GTPases activities and canonical YAP target genes. Meanwhile, administration of rLMO3 protein led to an opposite effect of LMO3 in the Hippo pathway.
The studies of Aoyama et al. and Isogai et al. showed that LMO3 interacts with HEN2 and enhances cell growth in neuroblastoma [14, 15]. Interestingly, in this research we found that LMO3 directly interacts with LATS1, and thus it suppressed Hippo signaling. Identifying the interaction between LMO3 and LATS1 provides direct evidence for the important role of LMO3 in the regulation of Hippo signaling. Also, we found that rLMO3 protein-induced HCC cell invasion and anoikis inhibition is abrogated by the inhibitors of the Hippo pathway, indicating that LMO3-induced HCC cell invasion, metastasis and anoikis inhibition are dependent on the suppression of Hippo signaling.
In conclusion, we found that LMO3 plays an important role in HCC cell invasion, metastasis and anoikis inhibition. High expression levels of LMO3 in HCC suppresses the Hippo signaling pathway by interacting with LATS1, and thus LMO3 promotes the invasion and metastasis of HCC cells. LMO3 may be used as a potential therapeutic strategy for HCC in future.
ANKRD1:
Ankyrin Repeat Domain 1
CNL:
Corresponding non-cancerous liver
CTGF:
Connective Tissue Growth Factor
CYR61:
Cysteine Rich Angiogenic Inducer 61
HCC:
LATS1:
Large Tumor Suppressor Kinase 1
LMO3:
LIM domain only 3
rLMO3:
recombinant LMO3
YAP:
Yes Associated Protein
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Zhang Z, et al. Migration of epithelial cells on laminins: RhoA antagonizes directionally persistent migration. Eur J Cell Biol. 2011;90:1–12.
Liu Y, Ding Y, Huang J, Wang S, Ni W, Guan J, et al. MiR-141 suppresses the migration and invasion of HCC cells by targeting Tiam1. PLoS One. 2014;9:e88393.
Wang YH, Dong YY, Wang WM, Xie XY, Wang ZM, Chen RX, et al. Vascular endothelial cells facilitated HCC invasion and metastasis through the Akt and NF-κB pathways induced by paracrine cytokines. J Exp Clin Cancer Res. 2013;32:51.
Chen B, Tang J, Guo YS, Li Y, Chen ZN, Jiang JL. Calpains are required for invasive and metastatic potentials of human HCC cells. Cell Biol Int. 2013;37:643–52.
Xie B, Xing R, Chen P, Gou Y, Li S, Xiao J, et al. Down-regulation of c-Met expression inhibits human HCC cells growth and invasion by RNA interference. J Surg Res. 2010;162:231–8.
Cui JF, Liu YK, Zhang LJ, Shen HL, Song HY, Dai Z, et al. Identification of metastasis candidate proteins among HCC cell lines by comparative proteome and biological function analysis of S100A4 in metastasis in vitro. Proteomics. 2006;6:5953–61.
Han Q, Lin X, Zhang X, Jiang G, Zhang Y, Miao Y, et al. WWC3 regulates the Wnt and Hippo pathways via Dishevelled proteins and large tumour suppressor 1, to suppress lung cancer invasion and metastasis. J Pathol. 2017;242:435–47.
Tong R, Yang B, Xiao H, Peng C, Hu W, Weng X, et al. KCTD11 inhibits growth and metastasis of hepatocellular carcinoma through activating Hippo signaling. Oncotarget. 2017;8:37717–29.
Varzavand A, Hacker W, Ma D, Gibson-Corley K, Hawayek M, Tayh OJ, et al. α3β1 Integrin Suppresses Prostate Cancer Metastasis via Regulation of the Hippo Pathway. Cancer Res. 2016;76:6577–87.
Wei C, Wang Y, Li X. The role of Hippo signal pathway in breast cancer metastasis. Onco Targets Ther. 2018;11:2185–93.
Salcedo Allende MT, Zeron-Medina J, Hernandez J, Macarulla T, Balsells J, Merino X, et al. Overexpression of Yes Associated Protein 1, an Independent Prognostic Marker in Patients With Pancreatic Ductal Adenocarcinoma, Correlated With Liver Metastasis and Poor Prognosis. Pancreas. 2017;46:913–20.
Qiao Y, Chen J, Lim YB, Finch-Edmondson ML, Seshachalam VP, Qin L, et al. YAP Regulates Actin Dynamics through ARHGAP29 and Promotes Metastasis. Cell Rep. 2017;19:1495–502.
Li C, Wang S, Xing Z, Lin A, Liang K, Song J, et al. A ROR1-HER3-lncRNA signalling axis modulates the Hippo-YAP pathway to regulate bone metastasis. Nat Cell Biol. 2017;19:106–19.
Nokin MJ, Durieux F, Peixoto P, Chiavarina B, Peulen O, Blomme A, et al. Methylglyoxal, a glycolysis side-product, induces Hsp90 glycation and YAP-mediated tumor growth and metastasis. Elife. 2016;19. https://doi.org/10.7554/eLife.19375.
Qiu YS, Jiang NN, Zhou Y, Yu KY, Gong HY, Liao GJ. LMO3 promotes gastric cancer cell invasion and proliferation through Akt-mTOR and Akt-GSK3β signaling. Int J Mol Med. 2018;41:2755–63.
This work was supported by Shanghai Municipal Science and Technology Commission (17401935000).
All data generated or analyzed during this research are included in this manuscript.
Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China
Yang Cheng
, Tianlu Hou
, Jian Ping
& Tianyang Chen
Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, 200040, China
Baobing Yin
Search for Yang Cheng in:
Search for Tianlu Hou in:
Search for Jian Ping in:
Search for Tianyang Chen in:
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BBY and YC designed the study. YC and TLH performed the study. JP and TYC participated in parts of the experiments and analysis of the data. BBY wrote the paper. All authors read and approved the final manuscript.
Correspondence to Baobing Yin.
All human samples were obtained with informed consent. The protocols were approved by the ethical review committee of the World Health Organization Collaborating Center for Research in Human Production (authorized by the Shanghai Municipal Government). All animals received humane care according to the criteria outlined in the "Guide for the Care and Use of Laboratory Animals" prepared by the National Academy of Sciences and published by the National Institutes of Health.
We have obtained consent to publish from the participant to report individual patient data.
Table S1 Primer sequences used for human LMO3, CTGF, ANKRD1 and CYR61 detection. a Edu assay of MHCC-97H cells infected with siRNA of LMO3 at 0, 24, 48 and 72 h time points respectively. b Edu assay of SMMC-7721 cells infected with siRNA of LMO3 at 0, 24, 48 and 72 h time points respectively. **P < 0.01. (DOC 30 kb)
Figure S2 The structure of pGreenPuro used for shRNA and vector construction. (PDF 622 kb)
Table S2 The original data of tissue microarray assay. (DOC 468 kb)
Figure S1 LMO3 knockdown has no effects on the proliferation of HCC cells. Multiplicity: 1 – single, 2 - multiple; Satelite: 0 – no, 1 or 2 - yes; Encapsulation: 0 – complete, 1 - incomplete; Vascular invasion: 0 – no, 1 - yes; Tumor thrombus: 0 – no, 1 - yes. (PDF 505 kb)
Cheng, Y., Hou, T., Ping, J. et al. LMO3 promotes hepatocellular carcinoma invasion, metastasis and anoikis inhibition by directly interacting with LATS1 and suppressing Hippo signaling. J Exp Clin Cancer Res 37, 228 (2018). https://doi.org/10.1186/s13046-018-0903-3
LMO3
Hippo signaling


Winner of the Black Cube Jam promoted by www.theblackcube.fr/ on Game Jolt, this short game, released in December 2016, was developed in 6 days only.
This remastered version contains some graphic improvements and additional contents, such as the full history of Riddoh.
Terran researchers have found evidence of a new undiscovered planet in the galaxy. Your journey begins landing on Riddoh.
Three strange domes tower over the city, now destroyed and uninhabited, as the last relics of an extinct alien civilization.
What secrets are contained in those domes?
Explore a mysterious land and solve some challenging puzzles.
If you'd like to give a small support and help us to continue developing other games we'd really appreciate it but, if you don't want to, just give our game an honest review and spread the word! Thanks!
For some reason, it won't download for me via the itch game app. When I click "install," it changes to "downloading," then a message pops up for about a microsecond (too fast for me to read it).
Ok, I disabled AdBlocker for the itch website, but it still won't download and install. I'm not sure what else I can do. Any suggestions?
Yeah I cannot install it either. I am very interested in the game though.
About the download, same old problem. If you use a pop-up blocker it prevent to open the download page.
Oddly enough, it worked just fine when I tried it again today without changing anything! Must have been a weird hiccup or something. Looking forward to checking it out!


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Quick Answer: How To Fight A Bus Stop Ticket?
Home > Bus ticket > Quick Answer: How To Fight A Bus Stop Ticket?
1 Is it illegal to block a bus stop?
2 How do you beat a bus lane ticket?
3 Can you park in front of a bus stop NYC?
4 How long is a NYC bus stop?
5 Can you legally stop someone parking outside your house?
6 Can you unload in a bus stop?
7 Do you get points for parking in a bus lane?
8 How do bus lanes work?
9 Is there an app to report illegal parking?
10 What do you do when someone doubles parks behind you?
11 What is illegally parked?
12 How many people can fit on a NYC bus?
13 What does no standing mean in New York?
Is it illegal to block a bus stop?
Stopping with any part of your vehicle (not just the wheels) on a restricted bus stop is likely to get you a hefty penalty charge. Rule 243 of The Highway Code tells road users: " Do not stop of park at or near a bus or tram stop or taxi rank".
How do you beat a bus lane ticket?
The simplest and most obvious way to avoid a bus lane ticket is to simply not drive or park one's car in a bus lane in New York City.
Can you park in front of a bus stop NYC?
You can beat a bus stop parking ticket if you are dropping off or picking up a passenger. But, try to avoid stopping temporarily in a bus stop zone. There must be at least one official NYC parking sign that designates the beginning of a bus stop zone, with an arrow indicating the direction(s) of the bus stop zone.
You might be interested: How Much Is A Greyhound One Way Bus Ticket From Seattle To Philadelphia?
How long is a NYC bus stop?
Thirty-two pairs of stops are less than 260 feet apart, or about the length of a city block. The MTA's guidelines say stops should be at least 750 feet apart, and international best practices put that number at 1,000 feet or more.
Can you legally stop someone parking outside your house?
Whilst there are no laws stopping people from parking directly outside your house, even if they do not live in your street, we can only hope for drivers to think about other people.
Can you unload in a bus stop?
Stopping to load and unload in a bus lane during its operational hours is only permitted so long as there are no loading or unloading restrictions in force. The use of bus lanes is strictly enforced and is restricted to buses and sometimes taxis and cycles.
Do you get points for parking in a bus lane?
Many bus lanes are monitored by CCTV, so the authorities should have a good understanding of your specific circumstances. But if you find you 've strayed into a bus lane by mistake, and you have no real excuse for doing so, you may be issued a penalty. You 'll be sent a penalty charge notice (PCN) through the post.
How do bus lanes work?
A bus lane is a lane restricted to buses on certain days and times, and generally used to speed up public transport that would be otherwise held up by traffic congestion. Bus lanes are shown by road markings and signs that indicate which (if any) other vehicles are permitted to use the bus lane.
You might be interested: FAQ: How To Excahnge Bus Ticket With Swiss Pass?
Is there an app to report illegal parking?
Snap Send Solve received over 45,000 reports of illegal parking in 2020. If you need to report an illegally parked car in your community, the Snap Send Solve app allows you to make a report in 30 seconds or less.
What do you do when someone doubles parks behind you?
You can call the police. Anything else you do, other than trying to find the owner of the car to move it, will be on you. If the car is illegally parked on your property, you may have other means of moving the car, such as calling a towing company.
What is illegally parked?
A parking violation is the act of parking a motor vehicle in a restricted place or for parking in an unauthorized manner. It is against the law virtually everywhere to park a vehicle in the middle of a highway or road; parking on one or both sides of a road, however, is commonly permitted.
How many people can fit on a NYC bus?
About the bus The bus provides approximately 42 percent more seating capacity, with 81 to 83 reclining seats, as opposed to 57 seats on standard express buses.
What does no standing mean in New York?
NO STANDING REGULATION. Sign indicates that vehicles may NOT stand at this. location. You may not wait or stop to load/unload. packages or merchandise at curbside.

redpajama_set_name: RedPajamaCommonCrawl


Our Silent Canvas – A non-profit multi-media arts collective presents ECHO THE ELEMENTS an evening of new works by four contemporary composers featured within a luminous forest installation created by our team of NYC based visual artists.
Each musical piece is in collaboration with various multi-media works including performance painting, interpretive dance, interactive electronics, and live VJ.
These twelve artists and composers offer several unique perspectives on the relationship between the rhythms of modern man and those of the natural world, exploring the sense of isolation or union that runs between them while encapsulating in their work the central theme ofEcho the Elements.
See more on OSC or Em.
← Cheat Sheet 5/9: LA Noire soundtrack, No Quarter, New Xbox?


The GET method is also used for more advanced job searches. Some field of the jobsummary descriptor can be used directly as parameters, and fields of the job descriptor can also be used as search criteria. You can also control the pagination and sorting order of the reply.
The name of the report job.
The username of the report job creator; the user who scheduled the report.
Gives the repository URI of a report to list all jobs. When this argument is omitted, this method returns all jobs for all reports.
Searches for jobs that match the JSON jobModel. The jobModel is a fragment of a job descriptor containing one or more fields to be matched.
Turns on pagination of the result by specifying the number of jobsummary descriptors per results page.
Determines the page number in paginated results by specifying the index of the first jobsummary to be returned.
Determines the sort order: ascending if true, descending if false or omitted.
200 OK – The body contains XML that describes jobs in the scheduler that match the search criteria.
The body of the return value is an XML jobs descriptor containing jobsummary descriptors, as shown in section Listing Report Jobs.
The example parameter lets you specify a search on fields in the job descriptor, such as output formats. Some fields may be specified in both the example parameter and in a dedicated parameter, for example label. In that case, the search specified in the example parameter takes precedence.


why do i crave ice?
Craving ice is often associated with iron deficiency anemia — although it may be associated with other nutritional problems as well.
Scientists don't fully understand the link, but some suspect that compulsive consumption of ice — called pagophagia — relieves inflammation in the mouth brought on by iron deficiencies.craving and chewing ice (pagophagia) is often associated with iron deficiency anemia — although it may be associated with other nutritional problems as well.
Make an appointment to have a complete blood test done to check for anemia.


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The 2 Top Stocks in Retail Apparel
Bradley Freeman, The Motley Fool
Here, we will explore two probable winners: Nike (NYSE: NKE) and Revolve Group (NYSE: RVLV). When considering iconic, globally ubiquitous brands, it's hard to overlook Nike. Over the past several decades the company has transformed itself as among the most valuable fashion brands in the world, and that is not by accident.
Ex-priest jailed for grooming and sexually abused three boys in 70s and 80s
Scottish fishermen turn on Boris Johnson over 'desperately poor' Brexit deal
Stephen Hardwicke, 63, was jailed for five years after being found guilty of abusing three boys - one as young as 10.
A top French general in West Africa has dismissed calls for his country to engage more in Central African Republic (CAR) after rebels earlier this week attempted to take the capital Bangui, saying that the situation was different to a rebellion in 2013. The Central African army has been battling groups backed by former president Francois Bozize that are seeking to overturn a Dec. 27 vote in which President Faustin-Archange Touadera was declared victor despite fraud claims. Russia and Rwanda have sent troops to back the government.
BEIJING — China on Saturday finished building a 1,500-room hospital for COVID-19 patients to fight a surge in infections the government said are harder to contain and that it blamed on infected people or goods from abroad. The hospital is one of six with a total of 6,500 rooms being built in Nangong, south of Beijing in Hebei province, the official Xinhua News Agency said. China had largely contained the coronavirus that first was detected in the central city of Wuhan in late 2019 but has suffered a surge of cases since December. A total of 645 people are being treated in Nangong and the Hebei provincial capital, Shijiazhuang, Xinhua said. A 3,000-room hospital is under construction in Shijiazhuang. Virus clusters also have been found in Beijing and the provinces of Heilongjiang and Liaoning in the northeast and Sichuan in the southwest. The latest infections spread unusually fast, the National Health Commission said. "It is harder to handle," a Commission statement said. "Community transmission already has happened when the epidemic is found, so it is difficult to prevent." The Commission blamed the latest cases on people or goods arriving from abroad. It blamed "abnormal management" and "inadequate protection of workers" involved in imports but gave no details. "They are all imported from abroad. It was caused by entry personnel or contaminated cold chain imported goods," said the statement. The Chinese government has suggested the disease might have originated abroad and publicized what it says is the discovery of the virus on imported food, mostly frozen fish, though foreign scientists are skeptical. Also Saturday, the city government of Beijing said travellers arriving in the Chinese capital from abroad would be required to undergo an additional week of "medical monitoring" after a 14-day quarantine but gave no details. Nationwide, the Health Commission reported 130 new confirmed cases in the 24 hours through midnight Friday. It said 90 of those were in Hebei. On Saturday, the Hebei government reported 32 additional cases since midnight, the Shanghai news outlet The Paper reported. In Shijiazhuang, authorities have finished construction of 1,000 rooms of the planned hospital, state TV said Saturday. Xinhua said all the facilities are due to be completed within a week. A similar program of rapid hospital construction was launched by the ruling Communist Party at the start of the outbreak last year in Wuhan. More than 10 million people in Shijiazhuang underwent virus tests by late Friday, Xinhua said, citing a deputy mayor, Meng Xianghong. It said 247 locally transmitted cases were found. Meanwhile, researchers sent by the World Health Organization were in Wuhan preparing to investigate the origins of the virus. The team, which arrived Thursday, was under a two-week quarantine but was due to talk with Chinese experts by video link. The team's arrival was held up for months by diplomatic wrangling that prompted a rare public complaint by the head of the WHO. That delay, and the secretive ruling party's orders to scientists not to talk publicly about the disease, have raised questions about whether Beijing might try to block discoveries that would hurt its self-proclaimed status as a leader in the anti-virus battle. Joe McDonald, The Associated Press
In a letter to the PM, the Scottish Fishermen's Federation (SFF) accused Johnson of misleading the public about the deal and giving the sector "the worst of both worlds."
EL TERRERO, Mexico — In the birthplace of Mexico's vigilante "self-defence" movement, a new group has emerged entirely made up of women, who carry assault rifles and post roadblocks to fend off what they say is a bloody incursion into the state of Michoacán by the violent Jalisco cartel. Some of the four dozen women warriors are pregnant; some carry their small children to the barricades with them. The rural area is traversed by dirt roads, through which they fear Jalisco gunmen could penetrate at a time when the homicide rate in Michoacán has spiked to levels not seen since 2013. Many of the women vigilantes in the hamlet of El Terrero have lost sons, brothers or fathers in the fighting. Eufresina Blanco Nava said her son Freddy Barrios, a 29-year old lime picker, was kidnapped by presumed Jalisco cartel gunmen in pickup trucks; she has never heard from him since. "They have disappeared a lot of people, a lot, and young girls, too," said Blanco Nava. One woman, who asked her name not be used because she has relatives in areas dominated by the Jalisco cartel, said that cartel kidnapped and disappeared her 14-year-old daughter, adding, "We are going to defend those we have left, the children we have left, with our lives." "We women are tired of seeing our children, our families disappear," the vigilante said. "They take our sons, they take our daughters, our relatives, our husbands." That is, in part, why the women are taking up arms; men are growing scarce in Michoacan's lime-growing hotlands. "As soon as they see a man who can carry a gun, they take him away," said the woman. "They disappear. We don't know if they have them (as recruits) or if they already killed them." Beside the barricades and roadblocks, the female vigilantes have a homemade tank, a heavy-duty pickup truck with steel plate armour welded on it. In other towns nearby, residents have dug trenches across roadways leading into neighbouring Jalisco state, to keep the attackers out. Alberto García, a male vigilante, has seen the medieval side of the war: He is from Naranjo de Chila, a town just across the river from El Terrero and the birthplace of the Jalisco cartel leader Nemesio Oseguera. Garcia said he was run out of the town by Jalisco cartel gunmen because he refused to join the group. "They killed one of my brothers, too," said Garcia. "They hacked him to pieces, and my sister-in-law, who was eight months pregnant." El Terrero has long been dominated by the New Michoacán Family and Viagras gangs, while the Jalisco cartel controls the south bank of the Rio Grande river. In 2019, the Viagras hijacked and burned a half-dozen trucks and buses to block the bridge over the river to prevent Jalisco convoys from entering in a surprise assault. And that same year, in the next town over, San Jose de Chila, the rival gangs used a church as an armed redoubt to fight off an offensive by Jalisco gunmen. Holed up in the church tower and along its roof, they tried to defend the town against the incursion, leaving the church filled with bullet holes. It is that stark divide where everyone is forced to chose sides — either Jalisco, or the New Michoacán Family and the Viagras — that has many convinced that the El Terrero vigilantes are just foot soldiers for one of those latter two gangs. The vigilantes bitterly deny allegations they're part of a criminal gang, though they clearly see the Jalisco cartel as their foe. They say they would be more than happy for police and soldiers to come in and do their jobs. El Terrero is not far from the town of La Ruana, where the real self-defence movement was launched in 2013 by lime grower Hipolito Mora. After successfully chasing out the Knights Templar cartel, Mora, like most of the original leaders, has distanced himself from the so-called self-defence groups that remain, and is now a candidate for governor. "I can almost assure you that they are not legitimate self-defence activists," said Mora. "They are organized crime. ... The few self-defence groups that exist have allowed themselves to be infiltrated; they are criminals disguised as self-defence." Michoacán's current governor, Silvano Aureoles, is more emphatic. "They are criminals, period. Now, to cloak themselves and protect their illegal activities, they call themselves self-defence groups, as if that were some passport for impunity." But in some ways, Mora says, the same conditions that gave rise to the original 2013 movement remain: Authorities and police fail to enforce the law and don't guarantee residents peace. Sergio Garcia, a male member of El Terrero vigilante group, says his 15-year-old brother was kidnapped and killed by Jalisco. Now, he wants justice that police have never given him. "We are here for a reason, to get justice by hook or by crook, because if we don't do it, nobody else will," Garcia said. ___ Associated Press writer Mark Stevenson contributed from Mexico City. Armando Solis, The Associated Press
WASHINGTON — President Donald Trump's impeachment trial is likely to start after Joe Biden's inauguration, and the Republican leader, Mitch McConnell, is telling senators their decision on whether to convict the outgoing president over the Capitol riot will be a "vote of conscience." The timing for the trial, the first of a president no longer in office, has not yet been set. But House Speaker Nancy Pelosi made it clear Friday that Democrats intend to move swiftly on President-elect Joe Biden's $1.9 trillion COVID aid and economic recovery package to speed up vaccinations and send Americans relief. Biden is set to take the oath of office Wednesday. Pelosi called the recovery package a "matter of complete urgency." The uncertainty of the scheduling, despite the House's swift impeachment of Trump just a week after the deadly Jan. 6 siege, reflects the fact that Democrats do not want the Senate trial proceedings to dominate the opening days of the Biden administration. With security on alert over the threat of more potential violence heading into the inauguration, the Senate is also moving quickly to prepare for confirming Biden's nominee for National Intelligence Director, Avril Haines. A committee hearing is set for the day before the inauguration, signalling a confirmation vote to install her in the position could come swiftly once the new president is in office. Many Democrats have pushed for an immediate impeachment trial to hold Trump accountable and prevent him from holding future office, and the proceedings could still begin by Inauguration Day. But others have urged a slower pace as the Senate considers Biden's Cabinet nominees and the newly Democratic-led Congress considers priorities like the coronavirus plan. Biden's incoming White House press secretary, Jen Psaki said Friday the Senate can do both. "The Senate can do its constitutional duty while continuing to conduct the business of the people," she said. Psaki noted that during Trump's first impeachment trial last year, the Senate continued to hold hearings each day. "There is some precedent," she said. Trump is the only president to be twice impeached, and the first to be prosecuted as he leaves the White House, an ever-more-extraordinary end to the defeated president's tenure. He was first impeached by the House in 2019 over his dealings with Ukraine, but the Senate voted in 2020 to acquit. When his second trial does begin, House impeachment managers say they will be making the case that Trump's incendiary rhetoric hours before the bloody attack on the Capitol was not isolated, but rather part of an escalating campaign to overturn the November election. It culminated, they will argue, in the Republican president's rally cry to "fight like hell" as Congress was tallying the Electoral College votes to confirm he'd lost to Biden. For Republican senators, the trial will be a perhaps final test of their loyalty to the defeated president and his legions of supporters in their states back home, and their own experiences sheltering at the Capitol as a pro-Trump mob ransacked the building and attempted to overturn Biden's election. It will force a further re-evaluation of their relationship with the defeated president, who lost not only the White House but majority control of the Senate. "These men weren't drunks who got rowdy — they were terrorists attacking this country's constitutionally-mandated transfer of power," said Sen. Ben Sasse, R-Neb., in a statement Friday. "They failed, but they came dangerously close to starting a bloody constitutional crisis. They must be prosecuted to the fullest extent of the law." McConnell, who has spent the past days talking to senators and donors, is telling them the decision on whether or not to convict Trump is theirs alone — meaning the leadership team will not work to hold senators in line one way or the other. Last week's assault angered lawmakers, stunned the nation and flashed unsettling imagery around the globe, the most serious breach of the Capitol since the War of 1812, and the worst by home-grown intruders. Pelosi told reporters on Friday that the nine House impeachment managers, who act as the prosecutors for the House, are working on taking the case to trial. "The only path to any reunification of this broken and divided country is by shining a light on the truth," said Rep. Madeleine Dean, D-Pa., who will serve as an impeachment manager. Trump was impeached Wednesday by the House on the single charge, incitement of insurrection, in lightning-quick proceedings just a week after after the siege. Ten Republicans joined all Democrats in the 232-197 vote to impeach, the most bipartisan modern presidential impeachment. McConnell is open to considering impeachment, having told associates he is done with Trump, but he has not signalled how he would vote. McConnell continues to hold great sway in his party, even though convening the trial next week could be among his last acts as majority leader as Democrats prepare to take control of the Senate with the seating of two new Democratic senators from Georgia. No president has ever been convicted in the Senate, and it would take a two-thirds vote against Trump, an extremely high hurdle. But conviction of Trump is not out of the realm of possibility, especially as corporations and wealthy political donors distance themselves from his brand of politics and the Republicans who stood by his attempt to overturn the election. Sen. Lisa Murkowski, R-Alaska, said Thursday, "Such unlawful actions cannot go without consequence." She said in a statement that the House responded "appropriately" with impeachment and she will consider the trial arguments. At least four Republican senators have publicly expressed concerns about Trump's actions, but others have signalled their preference to move on. Sen. Tom Cotton, R-Ark., issued a statement saying he opposes impeachment against a president who has left office. Trump ally Lindsey Graham of South Carolina is building support for launching a commission to investigate the siege as an alternative to conviction. The riot delayed the tally of Electoral College votes that was the last step in finalizing Biden's victory as lawmakers fled for shelter and police, guns drawn, barricaded the doors to the House chamber. A Capitol Police officer died from injuries suffered in the attack, and police shot and killed a woman. Three other people died in what authorities said were medical emergencies. ___ Associated Press writers Will Weissert, Kevin Freking, Andrew Taylor, Alan Fram, Zeke Miller and Jonathan Lemire contributed to this report. Lisa Mascaro And Mary Clare Jalonick, The Associated Press
WILMINGTON, Del. — President-elect Joe Biden is proposing a $1.9 trillion plan to expand coronavirus vaccinations, help individuals and jump-start the economy. The plan, which would require congressional approval, is packed with proposals on health care, education, labour and cybersecurity. On Friday, he outlined a five-step approach to getting the vaccination to the American people, and to ensure that it is distributed equitably. "Equity is central to our COVID response," he said. Here's a look at what's in Biden's plan: CONTAINING THE VIRUS — A $20 billion national program would establish community vaccination centres across the U.S. and send mobile units to remote communities. Medicaid patients would have their costs covered by the federal government, and the administration says it will take steps to ensure all people in the U.S. can receive the vaccine for free, regardless of their immigration status. — An additional $50 billion would expand testing efforts and help schools and governments implement routine testing. Other efforts would focus on developing better treatments for COVID-19 and improving efforts to identify and track new strains of the virus. THE VACCINATION PLAN — Working with states to open up vaccinations beyond health care workers, including to people 65 and older, as well as essential front-line workers. — Establishing more vaccination sites, including working with FEMA to set up 100 federally supported centres by the end of his first month in office . He suggested using community centres, school gymnasiums and sports stadiums. He also called for expanding the pool of those who can deliver the vaccine. — Using pharmacies around the country to administer the vaccine. The Trump administration already has entered into agreements with some large chains to do that. — Using the Defence Production Act, a Cold War-era law to "maximize the manufacture of vaccine and vaccine supplies for the country." — A public education campaign to address "vaccine hesitancy" and the refusal of some to take the vaccine. He called the education plan "a critical piece to account for a tragic reality of the disproportionate impact this virus has had on Black, Latino and Native American communities" INDIVIDUALS AND WORKERS — Stimulus checks of $1,400 per person in addition to the $600 checks Congress approved in December. By bringing payments to $2,000 — an amount Democrats previously called for — the administration says it will help families meet basic needs and support local businesses. — A temporary boost in unemployment benefits and a moratorium on evictions and foreclosures would be extended through September. — The federal minimum wage would be raised to $15 per hour from the current rate of $7.25 per hour. — An emergency measure requiring employers to provide paid sick leave would be reinstated. The administration is urging Congress to keep the requirement through Sept. 30 and expand it to federal employees. — The child care tax credit would be expanded for a year, to cover half the cost of child care up to $4,000 for one child and $8,000 for two or more for families making less than $125,000 a year. Families making between $125,000 and $400,000 would get a partial credit. — $15 billion in federal grants to help states subsidize child care for low-income families, along with a $25 billion fund to help child care centres in danger of closing. SCHOOLS — $130 billion for K-12 schools to help them reopen safely. The money is meant to help reach Biden's goal of having a majority of the nation's K-8 schools open within his first 100 days in the White House. Schools could use the funding to cover a variety of costs, including the purchase of masks and other protective equipment, upgrades to ventilation systems and staffing for school nurses. Schools would be expected to use the funding to help students who fell behind on academics during the pandemic, and on efforts to meet students' mental health needs. A portion of the funding would go to education equity grants to help with challenges caused by the pandemic. — Public colleges and universities would get $35 billion to cover pandemic-related expenses and to steer funding to students as emergency grants. An additional $5 billion would go to governors to support programs helping students who were hit hardest by the pandemic. SMALL BUSINESS — $15 billion in grants to more than 1 million small businesses that have been hit hard by the pandemic, as well as other assistance. STATE AND LOCAL GOVERNMENT — $350 billion in emergency funding for state, local and territorial governments to help front-line workers. — $20 billion in aid to public transit agencies. CYBERSECURITY — $9 billion to modernize information technology systems at federal agencies, motivated by recent cybersecurity attacks that penetrated multiple agencies. — $690 million to boost federal cybersecurity monitoring efforts and $200 million to hire hundreds of new cybersecurity experts. The Associated Press
Covid vaccination: 8100 to get jab on day one in Delhi
New Delhi [India], January 16 (ANI): Delhi Health Minister Satyendra Jain on Saturday said the vaccination drive has started at 81 centres in the national capital and 100 people would get their first of the two doses at each centre. Most of the Delhi government hospitals are serving as vaccination sites.
Warnings come into force as snowfall is seen across country
Margaret won £2650 and a spa break during Friday's episode of the game show
'Edge Of Tomorrow 2' will happen 'if Emily Blunt and Tom Cruise want it to'
Edge Of Tomorrow 2 could easily be made, if Emily Blunt and Tom Cruise decide that they want to star in it.
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If you do not have a strong password, you are susceptible to identity thefts and financial frauds.
3 Top Robinhood Stocks to Sell Before the Next Market Crash
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Yahoo Sport UK
Impatient O'Sullivan laments lack of practice as his silverware drought continues at Masters
Rusty Ronnie O'Sullivan slammed his lack of practice and admits he's itching to get his hands on silverware after crashing out of the Masters against old foe John Higgins, writes Will Jennings.
Matthew McConaughey reveals secret ambition to join the world of wrestling
Matthew McConaughey has teased that he might be about to join the WWE.
Germany: Armin Laschet elected leader of Merkel's party
BERLIN — Chancellor Angela Merkel's centre-right party has elected Armin Laschet, the pragmatic governor of Germany's most populous state, as its new leader — sending a signal of continuity months before an election in which voters will decide who becomes the new chancellor. Laschet defeated Friedrich Merz, a conservative and one-time Merkel rival, at an online convention Saturday of the Christian Democratic Union. Laschet won 521 votes to Merz's 466; a third candidate, prominent lawmaker Norbert Roettgen, was eliminated in a first round of voting. Saturday's vote isn't the final word on who will run as the centre-right candidate for chancellor in Germany's Sept. 26 election, but Laschet will either run for chancellor or will have a big say in who does. THIS IS A BREAKING NEWS UPDATE. AP's earlier story follows below. Conservative Friedrich Merz and pragmatic state governor Armin Laschet advanced to a runoff vote Saturday for the leadership of German Chancellor Angela Merkels's party, eight months before voters decide who will succeed Merkel in a national election. The new leader of the centre-right Christian Democratic Union will either run for chancellor in the Sept. 26 parliamentary election or have a big say in who does run. There had been no clear favourite going into Saturday's vote at an online convention of the CDU. A first round of voting among some 1,000 delegates produced a virtual tie between Merz and Laschet, with neither winning a majority. Merz, 65, is a one-time rival of Merkel who stands for a more conservative course but lacks government experience. He is making his second attempt to win the party leadership after a decade away from front-line politics, following his narrow loss in 2018 to outgoing leader Annegret Kramp-Karrenbauer. Merz told delegates that, when he joined the CDU at 16, "I didn't go into a placement agency for government jobs; I went into a party that has principles, that fought passionately for its politics ... always carried by the idea that there are no left-wing majorities in this country." Laschet, 59, is the governor of Germany's most populous state, North Rhine-Westphalia — a traditional centre-left stronghold where he won election in 2017. He is viewed as more in line with Merkel's pragmatic centrism, and pointed Saturday to the value of continuity and moderation. "We must speak clearly but not polarize," he told delegates. "We must be able to integrate, hold society together." Laschet said that "there are many people who find Angela Merkel good and only after that the CDU. He added that "we need this trust now as a party" and that "we must work for this trust." Merz won 385 votes and Laschet 380 in the first round of voting. A third candidate, Norbert Roettgen, was eliminated with 224 votes. After the runoff vote, Saturday's winner will be officially endorsed in a postal ballot — which is expected to be a formality but is required by German law. Saturday's decision will end an 11-month leadership limbo in the CDU after Kramp-Karrenbauer, who had failed to impose her authority on the party, announced her resignation. A vote on her successor was delayed twice because of the coronavirus pandemic. It won't be the final word on who runs to succeed Merkel as chancellor. The CDU is part of the Union bloc along with the Bavaria-only Christian Social Union, and the two parties will decide together on the centre-right candidate. The Union currently has a healthy poll lead, helped by positive reviews of Merkel's handling of the pandemic. CSU leader Markus Soeder, the governor of Bavaria, is widely considered a potential candidate after gaining in political stature during the pandemic. Some also consider Health Minister Jens Spahn, running to be the deputy CDU leader under Laschet, a possible contender. Merkel, who has been chancellor since 2005, announced in late 2018 that she wouldn't seek a fifth term. She also stepped down from the CDU leadership. Geir Moulson, The Associated Press
Azerbaijan will begin vaccinating citizens against COVID-19 on Monday, using a batch of 4 million doses from China's Sinovac Biotech Ltd, the health ministry said on Saturday. "Medical workers will be vaccinated first, and then over-65s from Feb. 1," presidential aide Shahmar Movsumov added. The doses will be transported first to Turkey, he said, where they will be checked and packaged, before arriving in batches to Azerbaijan.
Mohali (Punjab) [India], January 16 (ANI): Expressing happiness over the beginning of the vaccination drive against the novel coronavirus, Punjab Chief Minister Captain Amarinder Singh on Saturday said that there is no fear among the people about the vaccine.


But he can't out-fart Lin Carter.
Or he'd overload the vents.
Asimov himself has a valve let in his jeans.
He could shatter all the windows for miles around.
That could toot through Lily the Pink.
A stinker of a writer and a stinker when he farts.


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Mirando v. Mirando
Waterbury Trust Co. v. G. L. D. Realty Co.
Injunction is the usual and a proper remedy. Saratoga State Waters Corporation v. Pratt, 227 N.Y. 429, 447,…
Unifirst Corp. v. Rubino Bros., Inc.
"The law implies damage from the invasion of a legal right and recovery of nominal damages, at least, must be…
Full title:VITTORIA MIRANDO vs. JOSEPH MIRANDO
Court:Supreme Court of Connecticut Third Judicial District, New Haven, January Term, 1926
Date published: Apr 8, 1926
132 A. 910 (Conn. 1926)
132 A. 910
Supreme Court of Connecticut Third Judicial District, New Haven, January Term, 1926
In the comprehensive form of action known as ejectment or disseizin, a demand for possession and for damages may be joined with a prayer for equitable relief. The complaint in the present case examined and held to contain allegations sufficient to support a judgment as in an action of ejectment. The sum of $25 awarded a plaintiff who has offered no definite proof of loss will be regarded as the nominal damages which the law always implies from the invasion of a legal right. When the language of an instrument, clear upon its face, assumes a doubtful meaning in the light of the circumstances disclosed by the evidence, there is said to exist a latent ambiguity which is explainable by resort to extrinsic aid. The lease from the plaintiff to the defendant in the present case described the premises as a "garage" and a "store," the location and extent of which were, under the circumstances, doubtful. Held that evidence of the recent conduct of the parties, when joint owners of the premises, in dealing with a prior lessee was relevant in explanation of the ambiguity; and that its admission by the trial court was, in any event, harmless in view of the remaining testimony.
Argued January 27th, 1926
Decided April 8th, 1926.
ACTION in the nature of ejectment to recover possession of certain portions of premises owned by the plaintiff and occupied by the defendant under a claim that they were included in a lease to him by the plaintiff, brought to the District Court of Waterbury and tried to the court, Beardsley, Deputy-Judge; judgment for the plaintiff from which the defendant appealed. No error.
Frank P. McEvoy, for the appellant (defendant).
Charles G. Root, for the appellee (plaintiff).
HINMAN, J.
Plaintiff, who is the wife of the defendant but living apart from him, brings this action to recover from defendant possession of a cellar under a garage building, and a portion of a cellar containing a furnace and coal bin under the dwelling-house of plaintiff and back of a store; the garage and store having been leased to defendant by plaintiff, and he claiming possession, under the lease, of the cellar under the garage as a part of the garage, and of the portion of the cellar containing the furnace and coal bin as a part of the store. The parties entered into a lease which provided "that the lessor has leased and does hereby lease to the lessee the store 199 Hill Street and the garage in the rear." Since the lease, defendant has been in the occupancy of the store and the entire cellar in the rear thereof, and the garage building and cellar thereunder.
The court found that "the `store' leased by the plaintiff to the defendant, had been occupied jointly by the defendant and plaintiff before their separation, and included the whole of the basement within the four walls of the building, excepting the furnace, its enclosure and coal bin, and access thereto across the cement floor from the outside door of the building"; and that "the `garage' leased by the plaintiff to the defendant contained a two-car space facing Pilgrim Avenue and a one-car space facing Hill Street, but did not include the separate cellar underneath a portion of the `garage building' and unconnected therewith by entrance or by use." While these two paragraphs are stated in the finding as conclusions, they are subordinate facts.
The court held, among other conclusions, "that there was a latent ambiguity in said lease in the use of the words `store' and `garage,' explicable only by extrinsic evidence."
The defendant seeks, on appeal, to have the finding corrected by striking out six paragraphs. Of these, paragraphs ten and fourteen, and paragraphs one and two of subdivision II, are amply supported by evidence and by subordinate facts, set forth in the finding, which are based upon evidence and the court's inspection of the premises. Paragraph twenty-four is an obviously inadvertent and harmless repetition of a statement in a preceding paragraph of the finding. The conclusion stated in paragraph twenty, above quoted, is abundantly supported by subordinate facts. The remaining requests for correction were adequately complied with by the trial court.
The further reasons of appeal, with the exception of two hereinafter noticed, appear to be based upon a claim that the action as pleaded should be regarded as, in essence, one for reformation of the lease because of mutual mistake, and that, inasmuch as the finding negatives the existence of such mistake, the judgment rendered is not supported by the pleadings and is based upon facts which are not involved in the issues. Eliminating from consideration the paragraph alleging mistake, and the prayer for relief by reformation, the complaint yet contains allegations sufficient to that comprehensive form of action "indiscriminately called . . . ejectment or disseizin," which "lies for possessory rights," in which "damages may be recovered," and prayer for "equitable relief . . . may be joined with a demand for damages and possession." 1 Swift's Digest, p. 507; Walsh v. Feustel, 93 Conn. 366, 105 A. 696; Crandall v. Gallup, 12 Conn. 365. The judgment as rendered is appropriate to such an action, is within the prayers for relief, and is justified by the facts found.
The award of $25 damages without definite proof of loss or damage is also assigned as error. The law implies damage from the invasion of a legal right and recovery of nominal damages, at least, must be had. Beattie v. New York, N. H. H.R. Co., 84 Conn. 555, 80 A. 709; Dewire v. Hanley, 79 Conn. 454, 65 A. 573. The amount is not such as to render the damages awarded other than nominal. Brennan v. Berlin Iron Bridge Co., 72 Conn. 386, 44 A. 727.
The plaintiff had testified that, for a time before the defendant himself occupied the store, it was rented by one Mazaika, and had stated, without objection, that during his tenancy the door between the store and the cellar was locked. In reply to a question as to who occupied the cellar at that time, the plaintiff was permitted, over objection, to testify that she occupied it herself, and this ruling is assigned as error. The plaintiff then testified, without objection, that she kept wood, coal and groceries for family use in the cellar while Mazaika occupied the store. A latent ambiguity exists when, although particular words appear on the face of the instrument to be clear, it becomes necessary, in view of circumstances disclosed by the evidence, to resort to extrinsic aid in order to apply those words to the subject which they describe. Adams v. Turner, 73 Conn. 38, 46 A. 247; Petrie v. Trustees of Hamilton College, 158 N.Y. 458, 53 N.E. 216; Sherrod v. Battle, 154 N.C. 345, 70 S.E. 834; 3 Words Phrases (2d Series) p. 31; 2 Corpus Juris, 1314. Evidence of circumstances tending to show the meaning in which the words "store" and "garage" were used in the lease, and the subject-matter to which those terms related and its extent, was admissible. Brundrett v. Rosoff, 92 Conn. 698, 104 A. 67; Murphy v. Schwaner, 84 Conn. 420, 80 A. 295; Adams v. Turner, 73 Conn. 38, 46 A. 247; Hildreth v. Hartford, M. R. Tramway Co., 73 Conn. 631, 48 A. 963; Spongberg v. First National Bank, 15 Idaho, 671, 99 P. 712; Scheible v. Slagle, 89 Ind. 323. The recent conduct of the parties, when joint owners, in dealing with a stranger, was not irrelevant to the inquiry as to the meaning and scope of the word "store" as employed by them in their lease. Even if it could be held technically inadmissible, this isolated answer was harmless, in view of the other testimony to the same point, which preceded and followed it.


I have a friend who does videography, meaning videos of any kind, from slo-mo to nice sensitive ones. Constantin, by his name, has written a great blog post about 3 Ways To Get Paying Clients as a Videographer and he lives by what he preaches. See this video with a rail track in Brasov, Romania.
In the post I mentioned he explains a few things about the marketing a videographer needs to do. It's not all about the tech you have or your editing skills. Many clients will pay you by your marketing skills.
Constantin mentions a few great books in his post and among them 22 Immutable Laws of Marketing, a great book that should bring you up to speed with the basics in marketing. I just finished reading it and it gave me a birds-eye-view over my activities and some changes will result in the near future.
He mentions also three things you need to keep in mind if you want to get clients and the most intriguing is the first one: lie a bit, bend the truth. That is almost a requirement if you have no portfolio. If you believe in your skills invent something, then work on those and then go forward.


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The Negative Effects Of Dystopia In Ray Bradbury's Fahrenheit
Once there was a lonely man, his name unknown to the world. All his life he wondered what his purpose was. He dreamed of becoming a Broadway singer and actor, but he could not. His society restricted and made him work a job for the good of the society. They made him work as a handyman instead. As a result, he would be cursed with crippling depression because he could never do what he wanted. This brief story shows the true face of a dystopian society. Dystopias restrict freedoms and in the end leads to the breakdown of society. It is important to always be aware of the possible sign of a dystopia and stop them from happening, but sometimes no matter what people do their beloved world takes a turn for the worst. In the inspiring story of "Fahrenheit…show more content…
One of the main ways the author illustrates this is by subtly mentioning an ongoing war and a potential for a nuclear strike from an unknown source, the author says, "war may be declared any hour. This country stands ready to defend its… The firehouse trembled as a great flight of jet planes whistled a single note across the black morning sky" (Bradbury, 30). Throughout the story the main characters in the book rarely mention that there country is fighting an ongoing war and the possibility for a nuclear strike. This is because the characters want to feel safe even though their whole life could be taken away from them in the blink of an eye. The characters are too caught up in mindless entertainment that it becomes all they care about. The media brainwashes them into thinking they are safe because the people want to feel safe. This leads the characters to create a false sense of security. This makes the characters ignorant about situation within their country. This creates a dystopian world due to the overall lack of knowledge or care about their life. This further proves how the motif of fear best illustrates the negative effects of a dystopian society. Similarly, Bradbury continues to display the false sense of security when he talks about the need for firemen to burn houses that contain books, he states,…show more content…
The most important lesson one can learn from the powerful novel of Fahrenheit 451 is that in life people should always stay true to themselves no matter what. The minute people give up even the most basic freedom is a freedom they can never get back. In order to escape the disastrous effects of an unrelenting dystopia is to prevent them in first place and the only way to do this is to protect the basic freedoms people are blessed to have
Overcoming Successure Essay: How To Overcome Failure
OK, but what if we could take this fear of failure out of the question and attain the success that we want without unnecessary stress and too much trouble? Well, this is another thing. The famous published author Brian Tracy defined the fear of failure as the "single barrier between us and success" and we always use this "I can't" phrase to express this kind of fear. I can't do this, I can't do that… and we pretty much destroy our own success because we fear so much about not being able to do it, hence failing at life. Which leads me to ask myself: Is it something in the way we understand or define failure that makes us so fearful, so stressed up and in the end unsuccessful?
Schizophrenia Poem Analysis
The theme teaches you that you should do dirty things to other people if you don't want it to switch up and turn on you. If he had taken the time out to be patient and let his mom give it to him he would just have gotten a dime. In schizophrenia the mood was all based off anger between the couple, the house was just a bad environment for the both of them. The house crumbled and was tore apart physically, due to the lack of attention that it had from the couple in the house. The couple didn't have respect from each other or didn't act if they were appreciative to one another just as it was in the poem Those Winter Sundays.
Literary Analysis: The Giver By Lois Lowry
Jonas feel sad and misunderstood for the boy in war. Jonas sadly understood that no one know what he is feeling. These are like real life because some careless people don 't think about others and think that everything is just a joke. For example aborted a baby and people who play war games and don't care about real war that is happening on like the Syrian civil war. Besides in the novel Jonas realized that the community is living in a bad way and these
Analysis Of The Movie Marry Poppins
He hates banks and people who are working there. He considers that the whole world is guilty of his failures, but he believes he is not responsible for it. Travers Goff considers that he has no control over his life, and it depends only on other people. He says: "This world is just an illusion, Ginty, ol ' girl. As long as we hold that thought dear they can 't break us, they can 't make us endure their reality, bleak and bloody as it is.
Antigone: A Tragic Hero
These traits include the hero's tragic flaw, his position in society and his realization that his virtues had caused his demise. The tragic hero in Antigone is Creon, because he is a mature leader of society whose virtues (or flaws) cause his downfall. Creon is obdurate as he does not heed advice given from anyone during the majority of the play, he then finally follows the counsel that the Chorus Leader gives him near the end of the play. This is apparent during the argument between Haemon and Creon as Haemon tries to persuade him to listen to his subjects and change his opinions on the matter of Polyneices' burial as well as the incarceration of Antigone. Creon disagrees strongly and becomes inflamed towards Haemon.
Catcher In The Rye Depression Analysis
And further, when he does, he only ends up doing things that ruin the interaction with others, and makes himself become more isolated. Holden tries, but is always rejected and unsuccessful with his attempts. Holden is so desperate he manages to push people away even further. In the novel The Catcher In The Rye written by J. D. Salinger shows that Holden goes through his journey and is depressed because of his isolation from people and the world which he brings upon himself. As soon as the book starts, Holden isolates himself from the world and his friends.
Rhinotomy In Y Ha
In such a prosperous time, mournful music appears to be out of place; similarly, history seems to be outdated with the rapid development of the society. They can only move forward instead of looking back on history. Moreover, the onlookers' observation of the mad man's masochism is what truly makes his trauma an isolated business, thus the mad man is being alienated from the majority so as to bear the trauma all by himself. Their indifference to the torture that the mad man inflicted on himself indicates their apathy towards history.
Blood Diamond Criticism
First of all, we can see how the harshness of living conditions and the personal feelings can quickly demoralize a society as the situation cannot be controlled by anyone. The quest for personal power becomes the logic by which situations are judged and the ethical ramifications are often ignored all together. Next, leaders who operate under the guidance of darkness are quick to justify that their action are acting on behalf of those unable to make their own decisions. In reality, selfish interests often guide their decisions. In addition, a lack of morals and values respecting life and liberty, like those present in the RUF camps progresses to chaos and unethical actions.
Main Themes In Fahrenheit 451
(MIP-1) This meme showcases a central theme from the novel "Fahrenheit 451" by Ray Bradbury; the fact that books are feared and most of the people and the government perceive books as horrible, evil things. (SIP-1) Through this meme, readers can notice one of the reasons that books are feared, because they allow people to actually have feelings. (STEWE-1) This is brought to light when Faber tells Montag, "'So, now do you see why books are hated and feared? They show the pores in the face of life'" (79). Books are despised so much since the people of the society are so scared to go out of their comfort zone, which includes reading books.
Disadvantages Of Prostitution
Moreover, the employers follow a policy of isolation where they remove the documents and no one around them can speak their language except for their employer and they are also denied access to health care and other sources of assistance. The victims are fearful of the law enforcement agencies and therefore fail to recognize assistance as the victims are often told that if they come into contact with law enforcement, then their condition shall be worse than what is currently happening to them. There is a threat of exposure as the victims feel that they have failed and are ashamed from the society as result of which it results in them being trapped in similar kind of
More about The Negative Effects Of Dystopia In Ray Bradbury's Fahrenheit


What a crazy week it has been! A mix of (mostly) good and just a little bit bad, I'll try to catch up in photos. We decided to take the RV over to Gainesville for Mother's Day, sparing the kids the drive and the inconvenience. They all have such different schedules, it's really hard for all three to have a couple days off together. So, Sunday morning we loaded up the motorhome and made a leisurely drive across the state. Once again we stayed at Paynes Prairie.
We met Lindsey & Taylor for lunch at a wonderful local restaurant called Reggae Shack Cafe. There are so many vegan options here, it's hard to choose! I discovered on the last visit that they aren't messing around with their spice level, one of the dishes was too hot even for me heat-loving me! This time I choose the Tempeh Curry, which despite it's "brown-ness" was absolutely delicious. I ate way too much but I just couldn't help myself.
After lunch we hung out at the park for a little while and then met up with all of the kids at Kristyn's house. We were celebrating not only Mother's Day, but Jesus and Taylor's May birthdays as well. As always, Kristyn was the ultimate hostess! She cooked up an amazing vegan Manicotti, wish I'd taken a picture before gobbling it up! I did snap a quick shot of the cake, a (vegan, of course) marble cake w/salted caramel frosting. Oh, my goodness, so delicious! Provided by a local bakery, Sarkara Sweets, I'll definitely be ordering from them again!


The Week In Review is a weekly compilation of hand-picked NHL stories selected by members of WinColumnSports deemed important or impactful from the week that was. Additionally, there is also a running pool amongst the contributors based on our picks for Saturday's match-ups.
The Hart Memorial Trophy is awarded annually to the player deemed to be most valuable to his team through the regular season. It's a designation that has received a lot of debate over the years; does the award simply go to the best player in the league? Or does it go to a player that is so vital to his team's success that the team simply couldn't function without them?
If it's the latter, how does the trophy go to anyone other than Connor McDavid?
The Oilers are terrible when McDavid isn't on the ice; they've dropped from 8th to 21st in goals for, Cam Talbot's save percentage has dropped dramatically, and every defenceman not named Darnell Nurse has regressed this season. It's been a tire fire of a season in Edmonton after expectations were raised after last season.
However, that's no reason not to award the 21-year-old superstar his second consecutive Hart. He's invaluable to the Oilers franchise; while they may be a bad team now, imagine where the organization would be without him. He's third in league scoring, and the Oilers are a dramatically better team in both goals and Corsi for when he's on the ice.
If the Hart truly goes to the player most valuable to his team, is there really any argument?
Capitals goaltender Braden Holtby has been suspect this year to say the least.
Holtby is a 2x All-Star, Vezina Trophy winner in 2016 and was a huge part in the Capitals goal-tending duo winning the William M. Jennings award in 2017.
Holtby is struggling for the first time since being named the Capitals starter in 2012-2013 and a lot of it has to do with fatigue. Holtby is coming off his 3rd season of starting 63+ games, he started 72 games in 2014-2015.
Goalies don't have to be just physically prepared for every game, they have to be mentally prepared. Unlike other players who get to rest on the bench for 40 minutes a game, goaltenders have to be mentally engaged for the entire 60 minutes of the game.
As a Washington Capitals fan myself, I don't understand why we're forcing Holtby to carry such a big workload. We have a competent backup in Philipp Grubauer who is more than capable of playing 20-30 games a year.
Other goalies in the league are dealing with fatigue as well, questioning whether or not coaches should balance out the starts between goaltenders more. Just recently Andre Vasilevskiy of the league leading Tampa Bay Lightning made comments regarding fatigue.
Do you think goaltenders in the NHL carry too much of a workload?


Sat down and realized I was going to post a video of Eden… oopsie! That has to be started early in the day, and I fail.
Here's assorted photos from the mountain trail to make up for it… notice the height of the grass!!!
The short cowdog has to bounce to see where the cows are!
Long day in short glimpses!
The grass does look tall! You had a lot of helpers on this cattle drive. AndI think "Bounce" would be a cute name for a working ES.


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You are hereAbout Us / Staff / Jacquelin Green: Executive Director
Jacquelin Green: Executive Director
Jacquelin Green is our Executive Director, and coordinates all Outreach programs at YPSN. She has been working with families for over 20 years, and before that worked with people with HIV and in various outreach centres doing a whole lot of different things! Jacquelin feels really lucky to be able to work with parents and their little people both one-on-one and in groups. The strength, love and courage they show every day in building their families is really inspiring. She is also a mom to four terrific kids and knows just how challenging and wonderful being a parent can be.


Private label coffee pods made from an international blend of coffee beans and roasted in a BRC certified facility. Individual coffee pods are wrapped for freshness that work in most 1.0 and 2.0 single serve coffee makers (Kitchen-Aide®, Keurig® coffee pods, Bunn®, Breville®, Cusiniart® and others). Pooki's Mahi hazenut flavored single serve coffees is an intricately choreographed dance of flavor and balance that sweeps the palate away in a rush of symphonic delight.
Mocha Tasting Note(s):Medium roast coffee with hints of wine, chocolate, vanilla, macadamia nut and mocha.
Pooki's Mahi's new sustainable coffee pods, tea pods premiered at several Red Carpet Events and Celebrity Gift Bags.


DME Law School (Moot Court Committee), GGSIP University is pleased to announce the third edition of National Moot Court from 05th – 07th April 2019.
We look forward to the maximum participation of the students, all over the nation. Students participation is based on first come first serve basis.
The Registration form, rules and regulations, moot problem and the brochure are soon going to be uploaded. Kindly stay in touch to remain updated.


text:
Cain Accuser, Karen Kraushaar, Has History of Filing Workplace Complaints
In an AP Exclusive article, more light is being shed on Karen Kraushaar's past history of filing workplace complaints.
Three years after settling her complaint with Cain at the National Restaurant Association, Kraushaar, now a spokeswoman at the Immigration and Naturalization Service in the Justice Department, filed a complaint, with the assistance of her attorney, Joel Bennett, against her manager for circulating a sexually charged emailed in late 2002 or early 2003.
In order to settle the complaint, Kraushaar demanded thousands of dollars in compensation, a promotion on the federal pay scale as well as other items including a one-year fellowship to Harvard's Kennedy School of Government.
Later, she dropped the complaint, because it was "relatively minor".
Bennett has said that Kraushaar will appear in a joint news conference provided that the other women come forward. Neither Bennett nor Kraushaar have disclosed the exact nature and reason for the complaint against Cain.