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A Biased View of Bitcoin Account
Shes been home more than a week and never mentioned anything to me about it.I went into her room and looked under her pillow. It wasnt there. I looked under her bed, dragging out the storage boxes to find a better view, using my phone as a flashlight.Carla I asked.
Jane was in college, but I texted and requested her. She said she never saw an orange bit of paper.Wait, Carla said. We had the house cleaned while we were gone. Ill call them.Carla called the cleaning service married used and got the woman who cleaned the home on the line.
Nothing but egg cartons, espresso grinds, and Amazon boxes. The orange part of paper has been decomposing somewhere under a pile of garbage in a Los Angeles landfill.Carla requested if losing the newspaper was a big deal.Not actually, I said. Its a nuisance, thats all. Ill need to send all the bitcoins in the Trezor to an online wallet, reinitialize the Trezor, generate a new word list, and place the bitcoins straight back on the Trezor.
Its 551445. The Forgetting: April 4, 2017: 7.4 BTC $8,384I plugged the Trezor into my laptop and entered 551445. Wrong PIN entered.I must have made an error entering the PIN, I believed. I attempted 551445 again, taking good care to enter the digits correctly this time.Wrong PIN entered.Uh oh. I tried a small variation: 554445Wrong PIN entered.This is absurd, I thought.
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Id entered it at least a dozen occasions in recent months without having to refer to this paper. OK, its likely 554145. Wrong PIN entered.I looked at the tiny monochrome screen on the bitcoin wallet and noticed that a countdown timer had seemed. It was making me wait a couple of seconds before I could try out another PIN.
I went to the hardware pocket manufacturers website to learn about the PIN delay and also read the bad news: The delay doubled whenever a wrong PIN was entered. The website said, The number of PIN entry failures is saved in the Trezors memory. This means that power cycling the Trezor wont make the wait time proceed to zero again.
The thief would need to sit his life off entering the PINs. Meanwhile, you've got enough time to transfer your funds into a new device or wallet out of the newspaper backup. (Trezor is based in Prague, hence the stilted English.) The difficulty was, I was the thief, trying to sneak my own bitcoins back from my Trezor.
After my sixth incorrect PIN effort, creeping terror had escalated to heart-pounding panicI might have kissed my 7.4 bitcoins goodbye.I created a few more guesses, and each time I collapsed, my sense of unreality grew in proportion to the PIN delay, that was now 2,048 seconds, or about 34 minutes.
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One hundred guesses would take more than 80 sextillion years.I broke the news to Carla. I told her I couldnt remember the PIN and I was being punished every time I entered an incorrect PIN. She asked me if Id stored the PIN in my 1Password application (a secure password program ).
When she asked me why, I didnt have an answer.I knew it'd be find out this here a mistake you can try these out to waste a valuable guess in my agitated condition. My mind was becoming polluted with scrambled permutations of all PINs. I went into the kitchen to chop veggies for a curry we were making for dinner.
As I cut potatoes into cubes, I mentally shuffled around numbers like they were Scrabble tiles on a rack. After a time, a number popped into my head: 55144545. This was it! I walked from the kitchen to the workplace. The Trezor still had a couple hundred seconds left on the countdown timer.
I tapped in 55144545. Wrong PIN entered. Please wait 4,096 seconds to continueI barely slept that night. The small shuteye I managed to get has been filled with nightmares involving combinations of the numbers 1, 4, and 5. It wasnt so much the $8,000 that troubled meit was the shame I felt for being stupid enough to eliminate the paper and forget the PIN.
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When I wasnt able to recall the PIN, the Trezor would taunt me for the rest of my life.The Hunt: April 5, 2017: 7.4 BTC $8,325That morning, bleary eyed, I started looking into ways to receive my own bitcoins back that didnt involve recalling my PIN or recovery words. If Id lost my debit card PINI could contact my bank and Id eventually regain access to my funds.
No one owns the bitcoin transaction network. Instead, thousands of computers around the world run software that validates the systems transactions. Anyone is permitted to install the bitcoin software on their pc and participate. This decentralized nature of this bitcoin network is not without consequencesthe main one being that if you screw up, its your own damn problem.I went to /r/TREZOR/ on Reddit and posted:don't hesitate to ridicule meI deserve it.


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Original Research ARTICLE
MiR-409-3p Inhibits Cell Proliferation and Invasion of Osteosarcoma by Targeting Zinc-Finger E-Box-Binding Homeobox-1
- Minhang Hosptial, Fudan University, Shanghai, China
Osteosarcoma (OS) is the most common bone cancer worldwide. There is evidence that microRNA-409 (miR-409-3p) is involved in tumorigenesis and cancer progression, however, its possible role in OS requires clarification. In the present study, we evaluated the expression level, clinical significance, and mode of action of miR-409-3p in OS. The miR-409-3p levels were diminished in the OS cells and tissues compared with associated adjacent non-tumor tissues and a non-cancer osteoplastic cell line. Low miR-409-3p expression levels were associated with clinical stage and distant metastasis in patients with OS. Resumption of miR-409-3p expression attenuated OS cell proliferation and invasion. Additionally, based on informatics analyses, we predicted that zinc-finger E-box-binding homeobox-1 (ZEB1) is a possible target of miR-409-3p. This hypothesis was confirmed using luciferase reporter assays, reverse transcription-quantitative real-time polymerase chain reaction, and Western blot analyses. The findings of the current study indicated that ZEB1 was up-regulated in the OS tissues and cell lines, and that this up-regulation was inversely proportional to miR-409-3p expression levels. Furthermore, down-regulation of ZEB1 decreased OS cell invasion and proliferation, illustrating that the tumor suppressive role of miR-409-3p in OS cells may be exerted via negative regulation of ZEB1. Taken together, our observations highlight the potential role of miR-409-3p as a tumor suppressor in OS partially through down-regulation of ZEB1 and suggest that miR-409-3p has potential applications in OS treatment.
Osteogenic sarcoma (osteosarcoma; OS) is among the most common forms of bone cancer globally. The incidence ranges from 4 to 5 cases per million among children and teenagers (Gill et al., 2013; Tang et al., 2014). The OS tumors are always located in the distal femur or proximal tibia, and tumors in these regions present a high tendency to destroy adjacent normal tissues (Cates, 2016; Righi et al., 2016). Despite considerable advances in treatment strategies such as surgery, radiotherapy, chemotherapy and new antineoplastic agent (Li et al., 2015), cases with metastatic or recurrent OS have an inferior prognosis, and the likelihood of long-term survival for patients with advanced OS remains very low (Anderson, 2015; Isakoff et al., 2015). Genetic and epigenetic variations and potential environmental factors that block mesenchymal stem cell differentiation into osteoblasts contribute to OS tumorigenesis and tumor development (Sun et al., 2015; Li et al., 2018; Zhang et al., 2018), however, the detailed and complex molecular mechanisms underlying OS development remain largely unknown. Therefore, the molecular mechanisms underlying OS formation and progression require investigation to facilitate the development of novel therapeutic approaches for application in patients with OS.
MicroRNAs (miRNAs) are a subtype of endogenous, non-coding, single-stranded, short RNAs, with an approximate range in length of 19–25 nucleotides (Esteller, 2011). miRNAs can regulate the expression of protein-coding genes by binding to complementary sequences in the 3′-untranslated regions (3′ UTRs) of target genes, causing translational repression or mRNA cleavage (Bartel, 2004). miRNAs play key roles in various cellular processes, including apoptosis, cell proliferation, differentiation, angiogenesis, invasion, and metastasis (Gambari et al., 2016). Recently, the abnormal expression of miRNAs has been implicated in the etiology and development of various human cancers (Chen et al., 2016; Li and Wang, 2016; Wang et al., 2016). The potential biological roles of several miRNAs abnormally expressed in OS during its tumorigenesis have also been highlighted. For example, miR-422a expression is down-regulated in OS cell lines and tissues. Conversely, high levels of miR-422a expression can suppress OS cell invasion and proliferation, and improve paclitaxel and cisplatin-mediated apoptosis (Liu et al., 2016). Therefore, there is a need to explore the potential role of miRNA expression in OS and to unravel the underlying primary molecular mechanisms, which may provide information useful for designing new and efficient therapeutic strategies aimed at curing OS.
The effect of miR-409-3p has been investigated in various human malignancies, including breast (Ma et al., 2016), gastric (Zheng et al., 2012), colon (Tan et al., 2016), and prostate (Josson et al., 2015) cancers, however, its role of miR-409-3p in OS remains unclear. Latest study confirmed the interaction of miR-409-3p and ZEB1 played a role in the progression process of non-small cell lung cancer, indicating ZEB1 acted as a direct target of miR409-3p and could be modulated by miR-409-3p (Qu et al., 2018). Herein, we hypothesis there exists the miR-409-3p/ZEB1 axis in OS and report the first investigation of the expression levels, clinical significance, and biological functions of miR-409-3p in OS, as well as its underlying molecular mechanism.
Materials and Methods
Ethics Statement
All study participants voluntarily provided written consent before entering the study. We obtained the approval of The Ethics Committee of the Minhang Hospital, Zhongshan Hospital, Fudan University for Disease Control and Prevention. The methodology used in this study completely conformed to the recommendations of CONSORT 2010.
Tissue Specimens
Forty-nine pairs of osteosarcoma tumor and adjacent non-tumor tissues were collected from patients with osteosarcoma at Minhang Hospital, Zhongshan Hospital, Fudan University. No participants underwent chemotherapy or radiotherapy before surgery. All tissues samples were directly transferred into liquid nitrogen and were stored at -80°C until RNA extraction.
Cell Lines
OS cell lines, including HOS (GDC76) and MG63 (GDC074) were obtained directly from the Chinese Academy of Medical Sciences (Beijing, China) Cell Resource Center. A non-cancer osteoblastic cell line (hFOB 1.19 CRL-11372) was obtained from the American Type Culture Collection (ATCC; Manassas, VA, United States). All cells were incubated in Dulbecco's modified Eagle medium (DMEM; Gibco, Invitrogen Life Technologies, Carlsbad, CA, United States) supplemented with 10% fetal bovine serum (FBS; Gibco, Invitrogen Life Technologies, Carlsbad, CA, United States). All experimental cells were maintained at 37°C in 5% (V/V) carbon dioxide (CO2) and passaged every 2–3 days.
Cells were then seeded in 6-well plates at a density of 50–60% confluence for transfection. After overnight incubation, cells were transfected with miR-409-3p mimics, negative control miRNA mimics (miR-NC), ZEB1 siRNA, or scrambled siRNA (GenePharma, Shanghai, China), using Lipofectamine 2000 transfection reagent (Invitrogen, Carlsbad, CA, United States), according to the manufacturer's guidelines. Post-transfection (6 h), the culture medium was changed to DMEM containing 10% FBS.
Reverse Transcription-Quantitative Real-Time Polymerase Chain Reaction (RT–qPCR)
RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, United States), following the manufacturer's directions. A TaqMan Micro-RNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, United States) was used to reverse-transcribe miRNA, and qPCR performed using a TaqMan Micro-RNA PCR Kit (Applied Biosystems, Foster City, CA, United States). Reverse transcription of mRNA was performed using the M-MLV Reverse Transcription system (Promega Corporation, Madison, WI, United States). To determine ZEB1 mRNA expression levels we used the primers: forward 5′-AGGCAATAGGTTTTGAGGGCCAT-3′ and reverse 5′-TGCACCTTCTGTCTCGGTTTCTT-3′ and SYBR Premix Ex Taq (TaKaRa, Dalian, China). Endogenous U6 small nuclear RNA (primers: forward, 5′-CTCGCTTCGGCAGCACA-3′; reverse, 5′-AACGCTTCACGAATTTGCGT-3′) was amplified as an internal control for miR-409-3p, and β-actin (primers: forward, 5′-AGCGAGCATCCCCCAAAGTT-3′; reverse, 5′-GGGCACGAAGGCTCATCATT-3′) was amplified as an internal control for ZEB1 mRNA. All RT-qPCR experiments were conducted using an ABI7500 Real-time PCR system (Applied Biosystems, Carlsbad, CA, United States). Relative mRNA or miRNA expression levels were quantified using the 2-ΔΔCt method (Livak and Schmittgen, 2001).
3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) Assay
Post-transfection (24 h), cells were re-seeded into 96-well plates at 3,000 per well. Cells were maintained at 37°C in 5% (V/V) CO2 for 4 days. Then, cell proliferation was tested at the indicated times using the MTT assay (Sigma, St. Louis, MO, United States). In brief, 0.5 mg/mL MTT solution was added to cells, which were then incubated at 37°C for a further 4 h. Subsequently, we added 150.0 μL DMSO (Sigma, St. Louis, MO, United States) into each to dissolve the formazan crystals. Spectrometric absorbance was determined using a microplate reader (Bio-Rad Laboratories Inc., Hercules, CA, United States) at a wavelength of 490 nm.
Cell Invasion Assay
After 48 h transfection, cells were collected and suspended in FBS-free culture medium. Then, 5 × 104 cells were added into upper chambers of a 24-well Transwell Permeable Support device (8-μm pores, Costar; Corning Incorporated, Corning, NY, United States) coated with Matrigel (BD Biosciences, San Jose, CA, United States), while 500.0 μL culture medium containing 20% FBS was added to the lower chambers and cells incubated at 37°C in 5% CO2 for 48 h. We removed cells in the upper chambers using cotton swabs, then invaded cells were fixed with methanol, stained with 0.5% crystal violet, washed, and dried in air. An inverted microscope (Olympus Corporation, Tokyo, Japan) (200 × magnification) was used to calculate the number of invading cells in five randomly selected fields.
Prediction of miR-409-3p Targets and Luciferase Reporter Assays
Two miRNA targeted-gene databases, TargetScan1 and miRanda2, were used to predict target genes of miR-409-3p. HEK293T cells (ATCC) were seeded into 24-well plates at 40–50% confluence. After 24 h, cells were transfected with miR-409-3p mimics or miR-NC and pmirGLO-ZEB1-3′UTR-mutant (Mut) (GenePharma) or pmirGLO-ZEB1-3′UTR-wild-type (Wt) using Lipofectamine 2000. Cells were maintained at 37°C in 5% (V/V) CO2 for 48 h and luciferase reporter assays conducted using the Dual-Luciferase Reporter Assay System (Promega Corporation, Madison, WI, United States). Renilla luciferase was used as an internal control.
Western Blot Analyses
Cells were harvested after transfection for 72 h and lysed with RIPA Lysis Buffer (Beyotime Institute of Biotechnology, Haimen, China). Protein concentrations were determined using a BCA assay kit (PierceTM; Thermo Fisher Scientific, Inc.). Equal amounts of protein were separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes (Millipore, MA, United States), blocked with 5% skimmed milk for 2 h at room temperature, then incubated overnight at 4°C with mouse anti-human GAPDH monoclonal antibody (sc-137179; 1:1000 dilution; Santa Cruz Biotechnology) or mouse anti-human ZEB1 monoclonal antibody (sc-81428; 1:1000 dilution; Santa Cruz Biotechnology, CA, United States). Membranes were then washed three times using Tris-buffered saline containing 0.1% Tween-20 and probed with horseradish peroxidase-conjugated secondary immunoglobulin G goat anti-mouse (catalog no, sc-2005; 1:10,000) for 2 h at room temperature. Protein bands were visualized using enhanced chemiluminescence reagents (Bio-Rad Laboratories Inc., Hercules, CA, United States) and band densities analyzed using AlphaEase FC software (version 4.0.1; ProteinSimple, San Jose, CA, United States).
Statistical Analyses
Data were presented as means ± S.D. or box plots. We used SPSS 17.0 software (SPSS Inc., Chicago, IL, United States) for data analyses. Differences among groups were evaluated using one-way ANOVA corrected for multiple comparisons or Student's t-tests. The χ2-test was used to evaluate associations between miR-409-3p expression levels and clinicopathological factors. Spearman's correlation analysis was used to determine the correlation between miR-409-3p and ZEB1 mRNA expression levels. All statistical tests were two-sided; P < 0.05 were considered statistically significant.
MiR-409-3p Was Downregulated in OS Tissues and Cell Lines
RT-qPCR was used to evaluate miR-409-3p expression levels in OS tumor and adjacent non-tumor tissues. Expression of miR-409-3p was lower in OS tissues than that in adjacent non-tumor and normal tissue controls (Figure 1A, P < 0.05). Moreover, remarkable low levels of MiR-409-3p expression were detected in two OS cell lines relative to those in a non-cancer osteoblastic cell line (hFOB 1.19) (Figure 1B, P < 0.05).
Figure 1. Expression of miR-409-3p in OS tissues and cell lines. (A) Relative expression levels of miR-409-3p in 49 paired OS tumor and adjacent non-tumor tissues were evaluated by RT-qPCR. (B) Expression of miR-409-3p in OS cell lines compared with that in a non-cancer osteoblastic cell line (hFOB1.19). miR-409-3p, microRNA-409. OS, osteosarcoma. ∗P < 0.05, ∗∗P < 0.01 compared with the control group.
Relationship Between miR-409-3p Expression and OS Clinicopathological Factors
We also determined the relationship between miR-409-3p expression levels and OS clinicopathological factors. Our data showed that low miR-409-3p expression levels were significantly associated with advanced clinical stage (P = 0.035) and distant metastasis (P = 0.030), however, there were no significance associations with other clinicopathological factors, including sex (P = 0.961), age (P = 0.804), and tumor size (P = 0.851) (Table 1).
MiR-409-3p Reduces OS Cell Proliferation and Invasion
To investigate the role of miR-409-3p in OS, we transfected MG63 and HOS cells with miR-409-3p mimics, and used RT-qPCR to determine miR-409-3p expression levels (Figure 2A, P < 0.05). We investigated the role of miR-409-3p in OS cell proliferation using MTT assays conducted in MG63 and HOS cells transfected with miR-409-3p mimics or miR-NC. Expression of miR-409-3p led to a significant decline in MG63 and HOS cell proliferation (Figure 2B, P < 0.05). Similarly, the invasion capacity of HOS and MG63 cells transfected with miR-NC or miR-409-3p mimics was estimated using a cell invasion assay. As illustrated in Figure 2C, the introduction of miR-409-3p mimics into HOS and MG63 cells resulted in a considerable decline of invasion ability relative to the miR-NC group (P < 0.05). These observations suggested that miR-409-3p has a crucial role in the suppression of OS growth and metastasis.
Figure 2. The effects of miR-409-3p overexpression on cell proliferation and invasion in OS. (A) Relative expression of miR-409-3p in MG63 and HOS cells following transfection with miR-409-3p mimics or miR-NC. (B) MTT assays were performed to assess the effect of miR-409-3p overexpression on MG63 and HOS cells proliferation. (C) Cell invasion assays were conducted in MG63 and HOS cells following transfection with miR-409-3p mimics or miR-NC. miR-409, microRNA-409-3p (magnification, ×200). OS, osteosarcoma. miR-NC, negative control microRNA mimics. ∗P < 0.05, ∗∗P < 0.01 compared with the control group.
A Potential miR-409-3p Target in OS
We then investigated the molecular mechanisms underlying the tumor suppression caused by miR-409-3p in OS by predicting its potential targets using bioinformatics analysis. The 3′ UTR of ZEB1 was predicted to contain an miR-409-3p seed match at position 1280-1286 and has previously been reported as extensively upregulated in OS and participates in the regulation of OS tumorigenesis and progression (Shen et al., 2012; Li et al., 2016; Liu and Lin, 2016); therefore, we primarily focused on ZEB1 (Figure 3A) in this study. To validate the prediction, we performed luciferase reporter assays in HEK293T cells transfected with plasmids containing Mut and Wt ZEB 3′ UTR, along with miR-409-3p mimics or miR-NC. Luciferase activity was markedly downregulated in cells transfected with Wt ZEB1-3′ UTR and miR-409-3p mimics (Figure 3B, P < 0.01), however, no significant difference was observed in cells transfected with mutated ZEB1-3′ UTR and miR-409-3p mimics, suggesting that miR-409-3p could directly target the 3′ UTR of ZEB1. Additionally, RT-qPCR data showed that restoration of miR-409-3p expression led to down-regulation of ZEB1 mRNA expression in MG63 and HOS cells (Figure 3C, P < 0.01). Moreover, Western blot analysis demonstrated that miR-409-3p reduced ZEB1 protein expression in MG63 and HOS cells (Figure 3D, P < 0.05). In vivo assay showed the protein levels in tumor tissues were significantly lower than those in adjacent normal tissues (Figure 3E, P < 0.01). To summarize, Our data demonstratedthat ZEB1 is potentially a direct target gene of miR-409-3p in OS.
Figure 3. ZEB1 is a direct target of miR-409-3p in OS. (A) ZEB1 3′ UTR sequences containing wild type and mutant miR-409-3p binding sites. (B) Luciferase reporter assays performed in HEK293T cells co-transfected with miR-409-3p mimics or miR-NC, and pmirGLO-ZEB1-3′UTR Wt or pmirGLO-ZEB1-3′UTR Mut. After transfection (48 h), cells were collected and luciferase activities measured. ZEB1 mRNA (C) and protein (D) were detected in MG63 and HOS cells transfected with miR-409-3p mimics or miR-NC. miR-409, microRNA-409-3p. (E) Protein levels of ZEB1 in tumor tissues and adjacent normal tissues. OS, osteosarcoma. miR-NC, negative control microRNA mimics. Wt, wild type. Mut, mutant. ZEB1, Zinc-finger E-box-binding Homeobox-1. ∗P < 0.05, ∗∗P < 0.01 compared with the control group.
Upregulation of ZEB1 in OS Tissues and Negative Correlation of Its Expression With That of miR-409-3p
ZEB1 is recognized as a direct target gene of miR-409-3p in OS; therefore, we next investigated whether miR-409-3p expression levels were negatively correlated with those of ZEB1 in OS. Therefore, we performed RT-qPCR to evaluate ZEB1 mRNA expression levels and found that they were higher in OS specimens than adjacent non-tumor tissues (Figure 4A, P < 0.05). Moreover, Spearman's correlation analysis indicated an inverse relationship between miR-409-3p and ZEB1 mRNA expression (Figure 4B, r = -0.4725, P = 0.0006) in OS tissue samples. As shown in Figure 4C, we observed higher expression of ZEB1 mRNA and lower expression of miR-409-3p in tumor cell lines, when compared to normal cell line. Our results further confirm ZEB1 as a potential target of miR-409-3p in OS.
Figure 4. Inverse correlation between miR-409-3p and ZEB1 mRNA expression levels in OS tissues. (A) RT-qPCR analysis showing that ZEB1 mRNA levels were increased in OS tissues. (B) Spearman's correlation analysis of the association between miR-409-3p and ZEB1 mRNA in OS tissues. (C) Expressions of miR-409-3p and ZEB1 mRNA in cell lines. miR-409, microRNA-409-3p. OS, osteosarcoma. ZEB1, Zinc-finger E-box-binding Homeobox-1. mRNA, message RNA. ∗P < 0.05, ∗∗P < 0.01 compared with the control group.
Inhibition of ZEB1 Has Similar Effects to Those of miR-409-3p Overexpression in OS Cells
To explore the biological roles of ZEB1 in response to miR-409-3p inhibition in OS, we investigated whether ZEB1 knockdown mimicked the effects of miR-409-3p overexpression in OS cells. ZEB1-targeting siRNA was used to knockdown ZEB1 expression in HOS and MG63 cells. As shown in Figure 5A, ZEB1 protein was successfully knocked down in HOS and MG63 cells transfected with ZEB1 siRNA (P < 0.01). MTT and cell invasion assays showed that knockdown of ZEB1 by the introduction of ZEB1 siRNA suppressed MG63 and HOS cell proliferation (Figure 5B, P < 0.05) and invasion (Figure 5C, P < 0.05), suggesting that negative regulation of ZEB1 may mediate the tumor suppressive effects of miR-409-3p in OS cells.
Figure 5. ZEB1 under-expression has similar effects to miR-409-3p over-expression on OS cell proliferation and invasion. (A) ZEB1 protein expression was detected in MG63 and HOS cells transfected with ZEB1 siRNA or NC siRNA. MTT (B) and cell invasion (C) assays were conducted in MG63 and HOS cells transfected with ZEB1 siRNA or NC siRNA. miR-409, microRNA-409-3p (magnification, × 200). OS, osteosarcoma. ZEB1, Zinc-finger E-box-binding Homeobox-1. mRNA, message RNA. siRNA, small interfering RNA. NC, negative control. ∗P < 0.05, ∗∗P < 0.01 compared with the control group.
Dysregulation of miRNAs is a frequent event in various types of human cancer and has a pivotal role in the instigation of tumorigenesis and tumor progression where miRNAs can function as oncogenes or tumor suppressor genes (Fenger et al., 2014; Vanas et al., 2016). Furthermore, targeting miRNA with various types of chemically modified oligonucleotides has the potential to alter miRNA functions, providing a theoretical foundation for miRNA-based targeted therapy for specific human cancers (Trang et al., 2010; Imani et al., 2017; Tsai et al., 2017). Thus, research into the expression levels, biological roles, and fundamental molecular mechanisms of miRNAs has the potential to stimulate the development of novel approaches to the treatment of different types of cancer. Our data demonstrated that miR-409-3p expression levels were significantly down-regulated in OS tissues and cells relative to adjacent non-tumor tissues and a non-cancer osteoblastic cell line, respectively. Our observations are consistent with the findings of Ma et al. in breast cancer tissues and cell lines (Zheng et al., 2012). Additionally, reduced miR-409-3p expression levels were associated with clinical stage and distant metastasis in patients with OS, and our results also demonstrate that expression of miR-409-3p suppressed proliferation and invasion of OS cells. Furthermore, our data suggest that ZEB1 is a functional target of miR-409-3p in OS.
Recently, several studies have reported roles for abnormal miR-409-3p expression in the initiation and progression of various human cancers. For example, Josson et al. found that miR-409-3p expression was elevated in prostate cancer and that its re-expression in normal prostate fibroblasts resulted in a cancer-associated stroma-like phenotype, and miR-409-3p was released in extracellular vesicles to induce cancer initiation and epithelial-to-mesenchymal transition both in vitro and in vivo (Josson et al., 2015). Zheng et al. (2012) showed that miR-409-3p expression levels were decreased in gastric cancer and that they were negatively associated with tumor-node-metastasis stage and lymph node metastasis in patients with gastric cancer. Upregulation of miR-409-3p attenuated gastric cancer cell motility in vitro and decreased their ability to induce distal pulmonary metastases and peritoneal diffusion in vivo (Zheng et al., 2012). Tan et al. (2016) found that miR-409-3p was expressed at low levels in colon tumors and that its expression was negatively correlated with resistance to oxaliplatin. Ectopic expression of miR-409-3p improved the chemosensitivity of oxaliplatin-sensitive and oxaliplatin-resistant colon cancer cells (Tan et al., 2016). Therefore, miR-409-3p is a strong candidate for a new therapeutic target for the treatment of cancer because of its essential roles in cancer initiation and progression.
MiR-409-3p target identification is essential for understanding its potential functions in OS and developing novel targeted therapies for improving OS treatment. Potential miR-409-3p target genes have been previously reported; for example, Beclin-1 in colon cancer (Tan et al., 2016), radixin in gastric cancer (Zheng et al., 2012), and Ras suppressor 1 and stromal antigen 2 in prostate cancer (Josson et al., 2015). In the current study, we identified ZEB1 as a novel direct target of miR-409-3p in OS. Based on bioinformatics analysis, we predicted that ZEB1 contains a miR-409-3p seed match at position 1280–1286 of the ZEB1 3′ UTR. Luciferase reporter assays demonstrated that miR-409-3p directly targeted the 3′ UTR of ZEB1. Furthermore, Western blot and RT-qPCR analysis indicated that endogenous miR-409-3p has a negative regulatory effect on ZEB1 mRNA and protein expression in OS cells. Moreover, ZEB1 expression was high in OS tissues and inversely associated with that of miR-409-3p expression and knockdown of ZEB1 led to decreased OS cell proliferation and invasion, similar to miR-409-3p overexpression.
ZEB1, a member of the deltaEF1 family of two-handed zinc-finger transcription factors, maps to the short arm of human chromosome 10 (Zhang et al., 2019). ZEB1 expression is abnormally upregulated in various types of human cancer, including thyroid (Zhang et al., 2016), cervical (Ma et al., 2015), gastric (Jia et al., 2012), endometrial (Feng et al., 2014), and prostate (Drake et al., 2009) cancers. Accumulating evidence shows that ZEB1 has crucial roles during cancer initiation and progression (Kenney et al., 2011; Jia et al., 2012; Liu et al., 2012). In OS, Shen et al. reported that ZEB1 is highly expressed in tumor tissues and that its levels are significantly associated with lung metastasis. The signal network of ZEB1 involved in malignant transformation in various types of tumor is complicated. All of the upstream and downstream molecules participate in activating the signaling pathways in cell survival, senescence, chemosensitivity and immune escape, which may trigger the regulation of miR-409-3p. These findings suggest that inhibition of OS has the potential to be a novel and effective therapeutic target with the aim of curing this type of cancer. The limitations of this study include that we did not investigate the effects of ectopic ZEB1 over-expression on cell proliferation and invasion activity of miR-409-3p-expressing osteosarcoma cells and that the number of samples in this study is small thus multi-center trial is still needed.
In conclusion, here we establish for the first time that miR-409-3p expression is down-regulated in OS tissues and cell lines. Decreased miR-409-3p expression levels were associated with clinical stage and distant metastasis. MiR-409-3p targets ZEB1, which may be associated with OS carcinogenesis and progression, leading to inhibition of OS cell proliferation and invasion. Thus, the miR-409-3p/ZEB1 axis can be considered a novel therapeutic target for OS treatment. Further research is needed to explore whether the potential of miR-409-3p/ZEB1 can be realized to treat OS.
Author Contributions
These studies were conceived of and designed by all authors. Experiments were performed by LW and YZ. Data analysis, data interpretation, manuscript preparations were done by ZH, HG, KZ, XY, and JX.
The work was supported by General Projects of Shanghai Health State Planning Commission (No. 201640219).
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
We thank Australian Saqlain Abbas for helping us in editing our manuscript.
Anderson, M. E. (2015). Update on survival in osteosarcoma. Orthop. Clin. North Am. 2016, 283–292. doi: 10.1016/j.ocl.2015.08.022
Bartel, D. P. (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–297. doi: 10.1016/S0092-8674(04)00045-5
Cates, J. M. (2016). Utility of examination of biopsy tracts in osteosarcoma resection specimens. Am. J. Clin. Pathol. 146, 324–327. doi: 10.1093/ajcp/aqw119
Chen, X., Chen, X. G., Hu, X., Song, T., Ou, X., Zhang, C., et al. (2016). MiR-34a and miR-203 inhibit survivin expression to control cell proliferation and survival in human osteosarcoma cells. J. Cancer 7, 1057–1065. doi: 10.7150/jca.15061
Drake, J. M., Strohbehn, G., Bair, T. B., Moreland, J. G., and Henry, M. D. (2009). ZEB1 enhances transendothelial migration and represses the epithelial phenotype of prostate cancer cells. Mol. Biol. Cell 20, 2207–2217. doi: 10.1091/mbc.E08-10-1076
Esteller, M. (2011). Non-coding RNAs in human disease. Nat. Rev. Genet. 12, 861–874. doi: 10.1038/nrg3074
Feng, G., Wang, X., Cao, X., Shen, L., and Zhu, J. (2014). ZEB1 expression in endometrial biopsy predicts lymph node metastases in patient with endometrial cancer. Dis. Markers 2014:680361. doi: 10.1155/2014/680361
Fenger, J. M., Bear, M. D., Volinia, S., Lin, T. Y., Harrington, B. K., London, C. A., et al. (2014). Overexpression of miR-9 in mast cells is associated with invasive behavior and spontaneous metastasis. BMC Cancer 14:84. doi: 10.1186/1471-2407-14-84
Gambari, R., Brognara, E., and Spandidos, D. A. (2016). Targeting oncomiRNAs and mimicking tumor suppressor miRNAs: new trends in the development of miRNA therapeutic strategies in oncology (Review). Int. J. Oncol. 49, 5–32. doi: 10.3892/ijo.2016.3503
Gill, J., Ahluwalia, M. K., Geller, D., and Gorlick, R. (2013). New targets and approaches in osteosarcoma. Pharmacol. Ther. 137, 89–99. doi: 10.1016/j.pharmthera.2012.09.003
Imani, S., Wei, C., Cheng, J., Khan, M. A., Fu, S., Yang, L., et al. (2017). MicroRNA-34a targets epithelial to mesenchymal transition-inducing transcription factors (EMT-TFs) and inhibits breast cancer cell migration and invasion. Oncotarget 8, 21362–21379. doi: 10.18632/oncotarget.15214
Isakoff, M. S., Bielack, S. S., and Meltzer, P. (2015). Osteosarcoma: current treatment and a collaborative pathway to success. J. Clin. Oncol. 33, 3029–3035. doi: 10.1200/JCO.2014.59.4895
Jia, B., Liu, H., Kong, Q., and Li, B. (2012). Overexpression of ZEB1 associated with metastasis and invasion in patients with gastric carcinoma. Mol. Cell Biochem. 366, 223–229. doi: 10.1007/s11010-012-1299-6
Josson, S., Gururajan, M., Sung, S. Y., Hu, P., Shao, C., Zhau, H. E., et al. (2015). Stromal fibroblast-derived miR-409 promotes epithelial-to-mesenchymal transition and prostate tumorigenesis. Oncogene 34, 2690–2699. doi: 10.1038/onc.2014.212
Kenney, P. A., Wszolek, M. F., Rieger-Christ, K. M., Neto, B. S., Gould, J. J., Harty, N. J., et al. (2011). Novel ZEB1 expression in bladder tumorigenesis. BJU Int. 107, 656–663. doi: 10.1111/j.1464-410X.2010.09489.x
Li, C., Xu, W., Ding, J., Zhang, Y., Wang, J., Zhuang, X., et al. (2015). Micellization of antineoplastic agent to significantly upregulate efficacy and security. Macromol. Biosci. 15, 328–341. doi: 10.1002/mabi.201400356
Li, R. Z., and Wang, L. M. (2016). Decreased microRNA-452 expression and its prognostic significance in human osteosarcoma. World J. Surg. Oncol. 14:150. doi: 10.1186/s12957-016-0900-y
Li, S., Zhang, T., Xu, W., Ding, J., Yin, F., Xu, J., et al. (2018). Sarcoma-targeting peptide-decorated polypeptide nanogel intracellularly delivers shikonin for upregulated osteosarcoma necroptosis and diminished pulmonary metastasis. Theranostics 8, 1361–1375. doi: 10.7150/thno.18299
Li, Y., Zeng, C., Tu, M., Jiang, W., Dai, Z., Hu, Y., et al. (2016). MicroRNA-200b acts as a tumor suppressor in osteosarcoma via targeting ZEB1. Onco. Targets Ther. 9, 3101–3111. doi: 10.2147/OTT.S96561
Liu, C., and Lin, J. (2016). Long noncoding RNA ZEB1-AS1 acts as an oncogene in osteosarcoma by epigenetically activating ZEB1. Am. J. Transl. Res. 8, 4095–4105.
Liu, M., Xiusheng, H., Xiao, X., and Wang, Y. (2016). Overexpression of miR-422a inhibits cell proliferation and invasion, and enhances chemosensitivity in osteosarcoma cells. Oncol. Rep. 36, 3371–3378. doi: 10.3892/or.2016.5182
Liu, Z., Sun, B., Qi, L., Li, H., Gao, J., and Leng, X. (2012). Zinc finger E-box binding homeobox 1 promotes vasculogenic mimicry in colorectal cancer through induction of epithelial-to-mesenchymal transition. Cancer Sci. 103, 813–820. doi: 10.1111/j.1349-7006.2011.02199.x
Livak, K. J., and Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T) Method. Methods 25, 402–408. doi: 10.1006/meth.2001.1262
Ma, Y., Zheng, X., Zhou, J., Zhang, Y., and Chen, K. (2015). ZEB1 promotes the progression and metastasis of cervical squamous cell carcinoma via the promotion of epithelial-mesenchymal transition. Int. J. Clin. Exp. Pathol. 8, 11258–11267.
Ma, Z., Li, Y., Xu, J., Ren, Q., Yao, J., and Tian, X. (2016). MicroRNA-409-3p regulates cell invasion and metastasis by targeting ZEB1 in breast cancer. IUBMB Life 68, 394–402. doi: 10.1002/iub.1494
Qu, R., Chen, X., and Zhang, C. (2018). LncRNA ZEB1-AS1/miR-409-3p/ZEB1 feedback loop is involved in the progression of non-small cell lung cancer. Biochem. Biophys. Res. Commun. 507, 450–456. doi: 10.1016/j.bbrc.2018.11.059
Righi, A., Gambarotti, M., Sbaraglia, M., Sisto, A., Ferrari, S., Tos, A. P., et al. (2016). p16 expression as a prognostic and predictive marker in high grade localized osteosarcoma of the extremities: an analysis of 357 cases. Hum. Pathol. 58, 15–23. doi: 10.1016/j.humpath.2016.07.023
Shen, A., Zhang, Y., and Yang, H. (2012). Overexpression of ZEB1 relates to metastasis and invasion in osteosarcoma. J. Surg. Oncol. 105, 830–834. doi: 10.1002/jso.23012
Sun, L., Li, Y., Zhang, J., Li, H., Li, B., and Ye, Z. (2015). Prognostic value of pathologic fracture in patients with high grade localized osteosarcoma: a systemic review and meta-analysis of cohort studies. J. Orthop. Res. 33, 131–139. doi: 10.1002/jor.22734
Tan, S., Shi, H., Ba, M., Lin, S., Tang, H., Zeng, X., et al. (2016). miR-409-3p sensitizes colon cancer cells to oxaliplatin by inhibiting Beclin-1-mediated autophagy. Int. J. Mol. Med. 37, 1030–1038. doi: 10.3892/ijmm.2016.2492
Tang, J., Shen, L., Yang, Q., and Zhang, C. (2014). Overexpression of metadherin mediates metastasis of osteosarcoma by regulating epithelial-mesenchymal transition. Cell Prolif. 47, 427–434. doi: 10.1111/cpr.12129
Trang, P., Medina, P. P., Wiggins, J. F., Ruffino, L., Kelnar, K., Omotola, M., et al. (2010). Regression of murine lung tumors by the let-7 microRNA. Oncogene 29, 1580–1587. doi: 10.1038/onc.2009.445
Tsai, S. C., Lin, C. C., Shih, T. C., Tseng, R. J., Yu, M. C., Lin, Y. J., et al. (2017). The miR-200b-ZEB1 circuit regulates diverse stemness of human hepatocellular carcinoma. Mol. Carcinog. 56, 2035–2047. doi: 10.1002/mc.22657
Vanas, V., Haigl, B., Stockhammer, V., and Sutterluty-Fall, H. (2016). MicroRNA-21 increases proliferation and cisplatin sensitivity of osteosarcoma-derived cells. PLoS One 11:e0161023. doi: 10.1371/journal.pone.0161023
Wang, G. C., He, Q. Y., Tong, D. K., Wang, C. F., Liu, K., Ding, C., et al. (2016). MiR-367 negatively regulates apoptosis induced by adriamycin in osteosarcoma cells by targeting KLF4. J. Bone Oncol. 5, 51–56. doi: 10.1016/j.jbo.2016.02.002
Zhang, Y., Liu, G., Wu, S., Jiang, F., Xie, J., and Wang, Y. (2016). Zinc finger E-box-binding homeobox 1: its clinical significance and functional role in human thyroid cancer. Onco. Targets Ther. 9, 1303–1310. doi: 10.2147/OTT.S96723
Zhang, Y., Wang, F., Li, M., Yu, Z., Qi, R., Ding, J., et al. (2018). Self-Stabilized hyaluronate nanogel for intracellular codelivery of doxorubicin and cisplatin to osteosarcoma. Adv. Sci. 5:1700821. doi: 10.1002/advs.201700821
Zhang, Y., Xu, L., and Li, A. (2019). The roles of ZEB1 in tumorigenic progression and epigenetic modifications. Biomed. Pharmacother. 110, 400–408. doi: 10.1016/j.biopha.2018.11.112
Zheng, B., Liang, L., Huang, S., Zha, R., Liu, L., Jia, D., et al. (2012). MicroRNA-409 suppresses tumour cell invasion and metastasis by directly targeting radixin in gastric cancers. Oncogene 31, 4509–4516. doi: 10.1038/onc.2011.581
Keywords: osteosarcoma, microRNA-409, molecular mechanism, zinc-finger E-box-binding homeobox-1, invasion
Citation: Wu L, Zhang Y, Huang Z, Gu H, Zhou K, Yin X and Xu J (2019) MiR-409-3p Inhibits Cell Proliferation and Invasion of Osteosarcoma by Targeting Zinc-Finger E-Box-Binding Homeobox-1. Front. Pharmacol. 10:137. doi: 10.3389/fphar.2019.00137
Received: 22 October 2018; Accepted: 06 February 2019;
Published: 21 February 2019.
Edited by:Jian-ye Zhang, Guangzhou Medical University, China
Reviewed by:Jianmeng Wang, First Affiliated Hospital of Jilin University, China
Jixue Wang, Jilin University, China
Copyright © 2019 Wu, Zhang, Huang, Gu, Zhou, Yin and Xu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Jun Xu, email@example.com
†These authors have contributed equally to this work

subset: commoncrawl


text:
Awning Windows / August 2, 2018 / Olympia Doucet
Many new sliding windows have key locking latches already installed. Some allow the window to be locked slightly open for ventilation. If your home has these window locks they are usually secure enough to keep your insurance company happy. If your home has only a simple nonlocking latch you should secure them with a keylocking sliding window lock or a window grill. Installing window locks yourself will cost AUD$15$20 per window. Having these locks supplied and installed by a locksmith will cost $40$50 per window.
The kitchen sliding windows are made of two sashes that can slide horizontally over each other. There is a track that allows the left and right movement of the windows. You can add more that two sashes to make your windows taller and wider. Since the track that is made for sliding of the sashes is little above the ground it doesnt allow the dust to settle on it. This way it becomes easier to maintain the windows and dust will never jam the working of the windows thus increasing its life.
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Biomass Fuel Market Top Key Player Analysis By Diester Industries, Neste Oil Rotterdam, ADM, Infinita Renovables, Biopetrol, Cargill, Ital Green Oil, Glencore
This report researches the worldwide Biomass Fuel market size (value, capacity, production and consumption) in key regions like North America, Europe, Asia Pacific (China, Japan) and other regions.
This study categorizes the global Biomass Fuel breakdown data by manufacturers, region, type and application, also analyzes the market status, market share, growth rate, future trends, market drivers, opportunities and challenges, risks and entry barriers, sales channels, distributors and Porter's Five Forces Analysis.
Biomass Fuel Breakdown Data by Type
• Bioethanol
• Biodiesel
Biomass Fuel Breakdown Data by Application
• Industrial Fuels
• Transportation Fuels
• Chemical Industry
The Major players reported in the market include:
• Diester Industries
• Neste Oil Rotterdam
• Infinita Renovables
• Biopetrol
• Cargill
• Ital Green Oil
• Glencore
• Louis Dreyfus and More…
Click Here For More Details @ supplydemandmarketresearch.com/home/toc_publisher/42809?c...1
Objectives of this Report are:
• To analyze and research the global Biomass Fuel capacity, production, value, consumption, status and forecast;
• To focus on the key Biomass Fuel manufacturers and study the capacity, production, value, market share and development plans in next few years.
• To focuses on the global key manufacturers, to define, describe and analyze the market competition landscape, SWOT analysis.
• To define, describe and forecast the market by type, application and region.
• To analyze the global and key regions market potential and advantage, opportunity and challenge, restraints and risks.
• To identify significant trends and factors driving or inhibiting the market growth and More…
Table of Content:
Study Coverage
1.1 Biomass Fuel Product
1.2 Key Market Segments in This Study
1.3 Key Manufacturers Covered
1.4 Market by Type
1.4.1 Global Biomass Fuel Market Size Growth Rate by Type
1.4.2 Bioethanol
1.4.3 Biodiesel
1.5 Market by Application
1.5.1 Global Biomass Fuel Market Size Growth Rate by Application
1.5.2 Industrial Fuels
1.5.3 Transportation Fuels
1.5.4 Chemical Industry
1.6 Study Objectives
1.7 Years Considered
2 Executive Summary
2.1 Global Biomass Fuel Production
2.1.1 Global Biomass Fuel Revenue 2013-2025
2.1.2 Global Biomass Fuel Production 2013-2025
2.1.3 Global Biomass Fuel Capacity 2013-2025
2.1.4 Global Biomass Fuel Marketing Pricing and Trends
TOC Continued……!
Tables and Figures:
• Figure Biomass Fuel Product Picture
• Table Biomass Fuel Key Market Segments in This Study
• Table Key Manufacturers Biomass Fuel Covered in This Study
• Table Global Biomass Fuel Market Size Growth Rate by Type 2018-2025 (K MT) & (Million US$)
• Figure Global Biomass Fuel Production Market Share 2013-2025
• Figure Bioethanol Product Picture
• Table Major Manufacturers of Bioethanol
• Figure Biodiesel Product Picture
• Table Major Manufacturers of Biodiesel
• Table Global Biomass Fuel Market Size Growth Rate by Application 2018-2025 (K MT) and More…
About Us:
We have a strong network of high powered and experienced global consultants who have about 10+ years of experience in the specific industry to deliver quality research and analysis. Having such an experienced network, our services not only cater to the client who wants the basic reference of market numbers and related high growth areas in the demand side, but also we provide detailed and granular information using which the client can definitely plan the strategies with respect to both supply and demand side.
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Credit: Wochit Entertainment
Published on February 13, 2019 - Duration: 00:34s
Kim Kardashian Says The Trick To Wrinkle Free Face Is Not To Smile
Kim Kardashian attended an event held by her makeup artist Mario Dedivanovic.
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Category: Graduates
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Failing is not Failure You are not the only one to fail, you are not the first person to fail, you will not be the last person to fail; failing is not failure. Was JK […]
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If there is anyone element that you can change to make investing eight hours a day in an workplace simpler, its your chair. There is no shortage of evidence proving that becoming stuck inside a seat for too long can increase the risk of coronary disease and aggravate lower back pain. Actually, seated too much is even known as even worse than smoking. Many those who have workplace jobs develop problems like pins and needles, spine imbalance, joint pain, throat discomfort, and herniated dvds usually from seated too much time on the poor quality seat with out assistance. While you probably do not want to spend lots of money on the chair, a high-quality chair is an investment in your wellbeing, comfort and productivity. Two separate studies, released in 1990 and the year 2003, discovered efficiency goes up greater than 17% when folks operate in an ergonomic desk setting by having an flexible chair. The best chair combined with ergonomic training can also decrease place of work injuries. Even if you do not already have wrist or back issues, an ergonomic chair will help you keep up with the right position to prevent strain, carpel canal affliction, lower back pain, and spinal disc injury. The National Institute of Health recommends selecting a chair with all the required alterations in support proper posture. Including a seat with casters along with a 5-point foundation a seat skillet with thick, small-mobile froth padding or circles a backrest that is either curved or small enough to fit the small of the back soft armrests by having an adjustable peak and thickness a gas chair heat adjustment function and a tilt realignment to move someone weight towards the chairs backrest.


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2,229 Results Found
Beds 3 Baths 2 SqFt 2,375
MLS 884525 | Listing Courtesy of First Colorado Realty
Beds 2 Baths 3 SqFt 1,790
MLS 6874826 | Listing Courtesy of LoKation Real Estate
Beds 7 Baths 5 SqFt 7,003
MLS 8120079 | Listing Courtesy of Resident Realty
Beds 3 Baths 2 SqFt 1,950
MLS 8717606 | Listing Courtesy of Conscious Real Estate
Beds 3 Baths 2 SqFt 1,800
MLS 8304343 | Listing Courtesy of West and Main Homes Inc
Beds 3 Baths 4 SqFt 2,352
MLS 3101077 | Listing Courtesy of MB COLORADO REALTY NOW. INC
Beds 5 Baths 3 SqFt 2,325
MLS 5302391 | Listing Courtesy of HomeSmart Realty Group
Beds 3 Baths 2 SqFt 1,858
MLS 884523 | Listing Courtesy of Gateway Realty Colorado LLC
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Beds 3 Baths 2 SqFt 1,280
MLS 3934 | Listing Courtesy of Blue Tree Real Estate, LTD
Beds 5 Baths 2 SqFt 2,556
MLS 884521 | Listing Courtesy of Gateway Realty Colorado LLC
Beds 5 Baths 4 SqFt 4,100
MLS 884522 | Listing Courtesy of Shorewood Real Estate NOCO
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The content relating to real estate for sale in this Web site comes in part from the Internet Data eXchange (IDX) program of METROLIST, INC., DBA RECOLORADO®. Real estate listings held by brokers other than [insert IDX Subscriber's firm name here] are marked with the IDX Logo. This information is being provided for the consumers' personal, non-commercial use and may not be used for any other purpose. All information subject to change and should be independently verified.
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You have probably heard about the Wix web builder, when the company advertised their product during the 2015 SuperBowl games. As a publilcy traded company and market leaders, they aggressively advertise their product, neglecting the fact the the main product is free of charge. If the name Weebly rings a bell, it could be the fact that Tim Ferris, publisher of several best selling books and a top tier consultant recommends it with passion, stating it is one of the top website builders available, helping him build a fully functional web site in less than 2 hours. The rest are widely known as well, may it be for Ecommerce uses, or being a leading internet services company like Web, which is publicly traded at the NASDAQ stock exchange.
Support among the services varies widely, from free WordPress.com account's only offering community support, to Jimdo's email-only service, to Wix's telephone-callback service—even for free accounts! Many of the site builders offer rich online support knowledge bases and FAQs, so there's a good chance you won't even need to contact the company. I test each service's support as part of the review process by asking about some less-common site-building procedures.
I rarely comment on these sorts of reviews, but after reading your clearly unbiased and in depth review I felt it necessary to thank you. I already have a boldgrid website and domain and wanted to understand more about the limitations of that vs it's competitors, a LOT of other articles on the subjects are clearly shills for one of the companies, it's refreshing to see such an honest and thorough review, thanks again!
Social news A social news website features user-posted stories that are ranked based on popularity. Users can comment on these posts, and these comments may also be ranked. Since their emergence with the birth of web 2.0, these sites are used to link many types of information including news, humor, support, and discussion. Social news sites allegedly facilitate democratic participation on the web. Reddit, Digg, SlashDot
Free Website Builder USA
Some of the website builders will give you a free custom domain name for the first year if you sign up to their annual plans. After the first free year, it typically costs around $20 to renew every year. If you prefer to buy your own custom domain name elsewhere as it is usually cheaper, you can purchase your domain name is through GoDaddy, and it will cost you about $12 – $15 per year. From a bigger picture perspective, it's not a significant difference.
Internet Marketing Gurus List
In situations where you can't find an included tutorial to meet your needs, you can often find instruction from a variety of web resources. Whether you prefer written instructions or want to see each step in a video, you can find free information online to help you move forward with your design. So, before you think completing part of the setup of the site is too difficult, hope into your favorite search engine and see what if the larger community doesn't have an answer.
This guide is your training plan. As your trusty web host and coach, we're here to keep you on track as you build, grow, secure, and promote your new site. We've broken this website workout plan into goals and tasks to complete each quarter throughout 2019, but feel free to work at your own pace. You do the sweating, and we'll be right beside you with water cups and cheese-tastic motivational signs.
I'm pretty new to the whole web development/design aspect of things. I've tinkered before with free things but more specifically with forum design. I'm very interested in building a website but aside from having a main traditional website feel I'm looking to incorporate a forum to it. Would it be possible to do this with this WordPress/BlueHost tutorial here? Or would there be something you recommend for that sort of thing?


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~ Tullahoma TN topo map, 1:24000 scale, 7.5 X 7.5 Minute, Historical, 1972, updated 1988
Tullahoma, Tennessee, USGS topographic map dated 1972.
Includes geographic coordinates (latitude and longitude). This topographic map is suitable for hiking, camping, and exploring, or framing it as a wall map.
Printed on-demand using high resolution imagery, on heavy weight and acid free paper, or alternatively on a variety of synthetic materials.
Topos available on paper, Waterproof, Poly, or Tyvek. Usually shipping rolled, unless combined with other folded maps in one order.
- Product Number: USGS-5338070
- Free digital map download (high-resolution, GeoPDF): Tullahoma, Tennessee (file size: 14 MB)
- Map Size: please refer to the dimensions of the GeoPDF map above
- Weight (paper map): ca. 55 grams
- Map Type: POD USGS Topographic Map
- Map Series: HTMC
- Map Verison: Historical
- Cell ID: 45898
- Scan ID: 148741
- Imprint Year: 1988
- Woodland Tint: Yes
- Photo Revision Year: 1982
- Aerial Photo Year: 1979
- Edit Year: 1982
- Field Check Year: 1972
- Datum: NAD27
- Map Projection: Polyconic
- Map published by Tennessee Valley Authority
- Map Language: English
- Scanner Resolution: 600 dpi
- Map Cell Name: Tullahoma
- Grid size: 7.5 X 7.5 Minute
- Date on map: 1972
- Map Scale: 1:24000
- Geographical region: Tennessee, United States
Neighboring Maps:
All neighboring USGS topo maps are available for sale online at a variety of scales.
Spatial coverage:
Topo map Tullahoma, Tennessee, covers the geographical area associated the following places:
- Plainview Heights - Center Grove - Tullahoma - Westside Heights - Taylor Creek West Subdivision - Sharondale - Elkins - Pleasant Grove - Oak Park - Westwood - Johnstown - Forrest Park - Southgate Park - Pineview Heights - Bel-Aire - Estill Springs - Lakeside Estates - Taylor Creek East Subdivision - Kaywood
- Map Area ID: AREA35.37535.25-86.25-86.125
- Northwest corner Lat/Long code: USGSNW35.375-86.25
- Northeast corner Lat/Long code: USGSNE35.375-86.125
- Southwest corner Lat/Long code: USGSSW35.25-86.25
- Southeast corner Lat/Long code: USGSSE35.25-86.125
- Northern map edge Latitude: 35.375
- Southern map edge Latitude: 35.25
- Western map edge Longitude: -86.25
- Eastern map edge Longitude: -86.125
Related Maps
Breckenridge and Tennessee Pass, Colorado, Map 109 by National Geographic Maps
Chattanooga, Knoxville and Sevier County, Tennessee Street Map by Rand McNally
Chattanooga, Knoxville, and Sevier County, Tennessee by Rand McNally
Chattanooga, Tenneesee by The Seeger Map Company Inc.


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A British lawmaker revealed in Parliament on Thursday that he is HIV-positive.
Lloyd Russell-Moyle, a member of Parliament for the opposition Labour Party, is believed to be the only sitting lawmaker in the House of Commons to have disclosed that he has the virus.
"Next year, I'll be marking an anniversary of my own: 10 years since I became HIV-positive," the 32-year-old told the lower house. "It's been a long journey -- from the fear to acceptance and, from today, advocacy, knowing my treatment keeps me healthy and that it protects any partner I have."
Taking medication for a number of years has made him "what the [National Health Service] calls HIV-positive 'undetectable,' " he said.
Russell-Moyle, who was elected in 2017, explained that means he does not get sick and "can't transmit HIV to someone else."
"So as the virus lays undetectable and dormant in my body; my medication ensures that the virus doesn't reactivate, doesn't progress and can't be passed on. That's why the NHS says: Undetectable equals un-transmittable."
The MP told the Press Association news agency that he "had a duty as a Member of Parliament" to make the disclosure.
He said government cuts to sexual health budgets meant he could not "keep quiet" any more about an issue that "not only affects people I know, but it affects me so personally." The Department of Health and Social Care declined to comment.
Russell-Moyle added that he chose the timing of his announcement to mark the 30th World AIDS Day on December 1.
He is the second lawmaker to disclose that he is living with HIV. Former Labour Cabinet minister Chris Smith made the first revelation in 2005.
When someone is HIV-positive, regular treatment can nearly eliminate their chance of spreading the infection to others during sex.
Also Thursday, Public Health England reported that the number of people newly diagnosed with HIV in the UK fell 17% between 2016 and 2017.
An estimated 92% of the 102,000 people living with HIV in Britain have been diagnosed, the agency said. Of those, 98% are on treatment, and 97% of those on treatment are virally suppressed, meaning HIV is undetectable in their blood.
The figures make the UK one of the first countries to reach the UN's 90-90-90 targets: 90% of people with HIV will have a diagnosis by 2020, 90% of those will receive antiretrovirals, and 90% of those will have viral suppression.
About 39 million people were living with HIV/AIDS worldwide in 2017, according to UNAIDS, the United Nations program that sets international targets.
Have your say
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Thomas Dickson was born on 1 January 1891 in Wishaw, Lanarkshire. He was the third of eight children and the eldest son of Archibald Dickson and Elizabeth Stevenson. Archibald was the brother of Livingston Russell Dickson, my father's paternal grandfather. I have previously posted about Thomas' cousin, Archibald Dickson who was killed in France in … Continue reading Killed in Action- Thomas Dickson 1891-1917
A Young Soldier – Archibald Dickson 1898-1918
Archibald Dickson was my first cousin twice removed. His father and my father's maternal grandfather, Livingston Russell Dickson, were brothers. Archie, as he was known, was born in the family home at 216 Caledonian Road, Wishaw on 7 August 1898, the third son of coal miner, Robert Russell Dickson and Isabella Paterson. By 1901 the … Continue reading A Young Soldier – Archibald Dickson 1898-1918
David Russell Dickson 1864-1941
David Russell Dickson was my 2nd great uncle. His brother, Livingston was my paternal grandmother's father. I am writing about him today having stumbled over his obituary which gives me way more information about the man than I would find in the usual birth, death and marriage records. Starting from the beginning though, David was … Continue reading David Russell Dickson 1864-1941
Archibald Smith Rae 1903-1952
My paternal grandfather, Archibald Smith Rae, was born on 4 September 1903 in Craigneuk, Lanarkshire, Scotland. He was the third child of Robert Armstrong Rae, a coal minerand Margaret McGarrity. Robert and Margaret had six children between 1899 and 1925. Sarah Born 1899 John born 1901 Archibald Smith Born 1903 Robert Armstrong Born 1907 James … Continue reading Archibald Smith Rae 1903-1952


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Muhammad Zahidul Islam
Pathao downsizing on fund shortagePathao, a ride-hailing company and one of the fastest growing tech startups in Asia, cut more than 50 percent of its workforce in the last two days as it struggles to raise funds amid waning popularity for its services.
Shareholders may consider sale of Robi, says CEOThe government's move to raise the mobile operators' turnover tax to 2 percent from 0.75 percent from the new fiscal year will drive
Pay more for digital delightAt a time when the government should be encouraging virtual businesses, Finance Minister AHM Mustafa Kamal has proposed 7.5 percent value-added tax on them in his budget for fiscal 2019-20, making their services dearer.
Mobile phone calls may get costlierMobile phone services are likely to become costlier from today due to increased taxes. The National Board of Revenue sent a regulatory order to the mobile carries to increase the supplementary duty (SD) to 10 percent from 5 from midnight yesterday.
Robi to raise Tk 400cr thru' local bondsRobi is planning to raise Tk 400 crore from the local market by issuing a Shariah-based security bond with the face value of each certificate being Tk 1 lakh.


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When you are renting a 5, 10, 15, 20, 30 or 40 yard dumpster, you want a company you can trust with prices that make you smile. Give us a call today and see the difference we can make in your next construction or clean out project.
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Most cities or municipalities do not have many regulations regarding dumpster rental as long as you keep the dumpster entirely on your own property during the rental period. If you need to place your container on the street in any way, you will likely have to obtain a permit from the appropriate building permit office in your town. You can check with your local public works department to be sure.
Most dumpster rental companies will take care of securing this permit for you if you rent from them. Make sure that if you're planning to put the dumpster on the street, the company has made the appropriate arrangements. You should also ensure that you get the permit in a timely manner and at the correct cost. If you thought the dumpster company was getting a permit and they did not, you will be the one who will have to pay the fine that is issued by the authorities.
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At Mammoth Recycling we know that getting the proper dumpster in a well timed manner is an integral a part of making your project a success. Worth quotes for value to hire a dumpster in Guntersville typically comprise the following: the size of the container, the form of debris concerned, the base value for the dumpster, how a lot weight is contained within the quote, a given rental interval and delivery and pickup fees. If he were to dedicate himself to the activity as a full-time job, he says, discovering numerous discarded treasures, refurbishing and promoting them off, he is confident he might pull in no less than $250,000 a 12 months—there may be that a lot stuff simply tossed into dumpsters within the Austin space. There are haulage companies that also provide free waste consultation, helping companies in recognizing and applying essentially the most value-efficient inexperienced" constructing ways, conducting evaluation and waste audits for LEED reporting, while supplying correct waste disposal and recycling dumpsters. Worth quotes for hire a hire a dumpster in Chicago typically embrace the following: the size of the container, the kind of debris concerned, the base value for the dumpster, how a lot weight is contained within the estimate, a given rental interval and delivery and pickup fees. The rental firm may also have to know what date you want for delivery of the dumpster to your website as well as the variety of days you want it in order that the company can ensure they've the proper sized container obtainable for if you want it. Inventory could also be restricted at some rental companies, so it a good suggestion to call no less than a week ahead of time for worth quotes and scheduling delivering of a dumpster. The strategy works finest in case your angle of elevation is about 45o In different phrases that your distance from the tree is equivalent roughly to the height of the tree (you may get higher at estimating this, the more timber you measure). We will mechanically come to pick up the dumpster on the scheduled date or on the tenth day of the ten day rental interval. Subsequently, if the load rises over the top of the dumpster or overhangs, it's possible you'll be topic to extra charges. Name the development Rent a Construction Dumpster in Castaic provider if you're able to have it picked up. The container will then be weighed at a local landfill. Rental companies will commonly leave a roll off dumpster at your endeavor place for a lot of days or perhaps weeks. Eliminating gaps and making your stuff fit correctly can reduce the price of Rent a Construction Dumpster in Castaic. Because pianos are so heavy, it is likely that you will want a waste removal service to haul your old piano away. To get rid of Household Hazardous Waste please contact ERG Environmental Companies at 1-800-284-9107 or 419-354-6110. In case you have a flatbed you may as well pick up rental gear like forklifts and small to medium sized aerial gear to save lots of on delivery charges. Mixed recyclables and inexperienced waste must be sorted out to the blue and inexperienced carts, and the remaining refuse ought to be moderately compacted to make finest use of space. Name your neighborhood trash removal service at this time to schedule your convenient Rent a Construction Dumpster in Castaic, with time for spring simply. In case you are cleansing out your private home, doing some landscaping, planning a remodel , beginning a development project, or performing storm harm recovery , you'll likely find yourself in want of a Rent a Construction Dumpster in Castaic service. You can save time by placing every thing in your single stream trash container, and we'll take care of separating the recyclable materials and yard waste from the trash. So in case your trash hundreds are mild, however the Glad Tall Kitchen bag has trouble fitting in your trash can, the Gripper could be right for you. In fact it really is true that the dumpster rentals may harm your driveway or even correctly sorted when it arrives. In case you are renting a truck to take your waste to the landfill that value can quickly add up. To keep away from these costs, you possibly can simply let someone pick up your waste for you. Price can fluctuate vastly depending on what kind of service you need, as a brief dumpster is a one-time charge and a everlasting is an ongoing contract.
Dimensions: 12' Long x eight' Broad x 3' Tall Typical Uses: small scale reworking jobs, basement clean outs, garage clean ups and extra. One means to get actual estimates for the service you need would be to phone an area dumpster business and ask about their costs. So you wish to make sure that the dumpster you lease is appropriately sized for the job. Dumpsters are positioned in alleys and roll-out carts are available in residential areas with out alleys. The ten cubic yard Rent a Construction Dumpster in Castaic, CA is perfect for small cleanup jobs, inert waste and stable fill supplies comparable to broken concrete, asphalt, block, grime, brick. With the meter below the floor, the opening at the meter may develop into submerged in a non-potable (not secure for ingesting) liquid and allow contamination to enter the water system. Dumpster rental corporations usually have set costs, however the high-quality artwork of negotiation is consistently in play. 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I can not tell you what number of occasions individuals have informed me that the food I get out of the dumpster is more energizing than what they've bought at the store. Even when the world where the dumpster is going to be positioned is paved, it's good apply to secure plywood to tempo under the unit's wheels to assist keep away from injury to the property. The only reason Waste Management gets jobs is as a result of it is a family identify and it's a first cease for many people who need leases. While you call to get a price quote, have really useful of simply how much waste you may must take away so you could get the best suggestion on dumpster measurement. Covered by your stable waste companies fee ($11/month/unit), collected quarterly on your water invoice. If in any respect potential, be available for delivery, so you'll be able to information the dumpster delivery driver in the placement of the container. There are usually two forms of dumpster you'll be able to lease primarily based on the type of waste that the dumpster is made for - one for property or family particles and one for food and basic blended wastes that embrace extra natural supplies. Looking for the nearest Rent a Construction Dumpster in Castaic, CA in your space from the listing or the net. The original price was affordable, however the total price was nearly double the amount. DTM accurately models the ground floor and permits the estimator to straight calculate volumes with out drawing counters. Both approach, please remember that the contracted digital recycling firm cleans the exhausting drives of all forms of computer systems with Division of Protection approved software program. Most how much does a dumpster price to lease corporations in Mattawa usually lease containers for a normal price for 7 days, although most one time clients find yourself conserving their container a median of three and a half days. Relying on the fees at the landfill, your costs might be substantially completely different than in other areas. Please contact your community supervisor in case you have bulk gadgets comparable to furnishings pieces, or boxes of outdated kitchen & home goods, and so on. We provide residential trash and recycling assortment to homes in many areas of Connecticut. There are plenty of things to think about when deciding on an area or national how much does it price to lease a dumpster agency in Goodwater. Must you believe you may need the dumpster for an additional week or extra, make sure that to mention that; charges may change primarily based on an extended time period. Relying upon the fees at the landfill, your costs could be considerably completely different than in other cities. The 30-Yard dumpster is perfect for new construction, demolitions, estate clean-outs and landscaping particles removal. While you lease a roll-off dumpster, you may be enticed to throw anything and all the pieces indoors. To keep away from rodents and other pest infesting your space, renters can get dumpsters with tightly fitted lids and doors and make sure the openings are closed always; inserting the dumpster on concrete or asphalt instead of grime is an effective way to stop rodents from burrowing underneath.
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