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Petkov SG1, Glage S2, Niemann H3Author Names in full: Stoyan G Petkov1, Silke Glage2, Heiner Niemann3

1 German Primate Center, Goettingen, Germany, 2 Hannover Medical School, Hannover, Germany, 3 Institute for Farm Animal Genetics (FLI), Neustadt, Germany
doi:10.46582/jsrm.1301004
Citation: Petkov SG, Glage S, Niemann H3. Mouse iPSC generated with porcine reprogramming factors as a model for studying the effects of non-silenced heterologous transgenes on pluripotency. J Stem Cells Regen Med 2017; 13(1) :20-28. doi:10.46582/jsrm.1301004 Abstract
Mouse somatic cells can be reprogrammed to pluripotency by the ectopic expression of four pluripotency transcription factors, Oct4, Sox2, c-myc, and Klf4. Usually, silencing of the exogenous reprogramming factors is considered to be essential for complete reprogramming and differentiation. In the vast majority of studies, murine pluripotency transcription factor sequences have been used for the reprogramming of mouse fibroblasts to induced pluripotent stem cells (iPSC). The effectiveness of xenogeneic transcription factors in miPSC generation has not yet been investigated in detail. Here, we evaluated transposon-based vectors with four porcine pluripotency factors for their ability to reprogram mouse fetal fibroblasts (MEFs) harboring an Oct4-EGFP reporter construct to pluripotency. Additionally, we examined the effects of the non-silenced heterologous transgenes on the expression levels of key endogenous pluripotency markers and the differentiation capacities of the miPSC. Within 8 days of transfection with porcine reprogramming transcription factors the MEFs acquired typical compact miPSC morphology and upregulated expression of endogenous Oct4 and other critical pluripotency genes. Consequently, the transgenes under the control of the TetO promoter became silenced, while the CAG-controlled constructs were expressed throughout the period of culture. Despite the continuous transgene expression, the CAG-miPSC showed normal morphology and were capable of differentiation into the three primary germ layers in vitro and in vivo. However, the expression levels of important endogenous pluripotency markers, Klf4, c-myc, Rex1, and Utf1, were significantly lower in CAG-miPSC compared with TetO-miPSC with silenced reprogramming cassettes. Surprisingly, the endogenous Oct4 and Sox2 expression levels were not affected by the residual transgene expression. Our results suggest that porcine reprogramming transcription factors are suitable for production of miPSC, but silencing of the heterologous transgenes may be necessary for complete reprogramming to pluripotency.
Keywords: iPSC; Reprogramming; Silencing
Introduction
The fate of a terminally differentiated somatic cell can be reversed by ectopic expression of pluripotency-related factors, as demonstrated by Takahashi and Yamanaka[1]. In experiments that have since revolutionized the stem cell field, it was demonstrated that elevated expression of only four proteins such as Oct4, Sox2, c-myc, and Klf4 leads to upregulation of endogenous pluripotency transcription factors and ultimately to acquisition of pluripotent characteristics by the reprogrammed cells[2,3]. To underscore the difference to alternative reprogramming methods (e.g. by somatic cell nuclear transfer, cell extracts, etc.), the resulting stem cells have been termed “induced pluripotent stem cells” (iPSC). During the reprogramming process, the balance between the expression levels of the endogenous pluripotency genes and the transgenes is thought to be critical for the success of the reprogramming of somatic cells to iPSC, with the epigenetic silencing of the retroviral transgenes coinciding with the up-regulation of the endogenes[4]. It has been shown that over-expression of Oct4 in mouse embryonic stem cells (mESC) leads to mesodermal differentiation[5], while elevated Sox2 expression causes neuroectodermal specification[6]. Consequently, the net concentration levels of Oct4 and Sox2 in the iPSC would need to be maintained within a narrow range characteristic for ESC. In support of this hypothesis, it was demonstrated that following upregulation of the corresponding endogenous pluripotency factors, the transgenes became gradually silenced in successfully reprogrammed iPS cells[7,8]. Persisting transgene expression results in perturbations in the reprogramming process, usually leading to partially reprogrammed states (Class I iPSC), as shown by Mikkelsen and co-workers[9]. Residual transgene expression restricted the differentiation capacities of human iPSC[10] and affected epigenetic signatures and gene expression levels of miPSC compared with mESC[11]. Moreover, in a recent report, miPSC produced with transposon-delivered transgenes that were continuously expressed by supplementation with doxycycline (DOX) were reversed to an alternative pluripotency state (F-state), characterized by aberrant colony morphology and dependency on the continuous transgene expression for their maintenance[12].
It has been convincingly demonstrated that the differentiation capacities of murine iPSC lines with constitutive expression of non-silenced reprogramming factors are limited. However, conflicting results were reported by different research groups. The miPSC produced in the first attempts of Takahashi and Yamanaka[1] did not silence the reprogramming factors and were not able to differentiate and form chimeras. Similarly, transgene-expressing miPSC were unable to differentiate and upon injection into immunodefficient mice formed homogenous tumors, consisting of undifferentiated cells expressing high levels of Oct4[13]. On the other hand, the use of a constitutively expressed reprogramming cassette led to miPSC with normal ESC-like characteristics, which were capable of forming the three primary germ layers in teratomas[14]. However, these cells also exhibited gene expression differences compared with ESC and excision of the loxP-flanked transgenes resulted in miPSC with complete pluripotency characteristics.
While the vast majority of miPSC described in the literature have been generated with murine pluripotency transcription factors, there have been very few reports on the use of transgenes from other species. Only one study has reported the use of human reprogramming factors to successfully reprogram mouse fibroblasts to iPSC[15]. In addition, the silencing of non-murine reprogramming factors as well as the effects of the non-silenced xenogeneic transgenes on the expression of endogenous pluripotency genes and its relationship with differentiation of miPSC have not been yet investigated. Here, we explored the possibility to use porcine transcription factors for the reprogramming of mouse embryonic fibroblasts (MEFs) to pluripotency. Established miPSC lines constitutively expressing the four porcine Yamanaka factors (pOCT4, pSOX2, pc-MYC, and pKLF4) under the control of the ubiquitous CAG promoter were characterized for pluripotency and their differentiation potential together with the expression levels of master pluripotency genes were compared with those of miPSC produced either with a silenced or CRE recombinase-excised reprogramming cassette.
Materials and methods
Unless specified otherwise, the chemicals and kits used in this study were purchased from Sigma-Andrich.
Derivation of mouse fetal fibroblasts for feeder cells and reprogramming
Outbred NMRI mice were purchased from Harlan Laboratories (Horst, Netherlands) and were used to establish MEFs for feeder layers. C57BL/6 mice harboring Oct4-EGFP pluripotency gene reporter (OG2) were kindly provided by Prof. Hans Schoeler (Max-Planck-Institute for Molecular Biomedicine, Muenster, Germany) and were used to generate MEFs for reprogramming to iPSC. For the generation of MEFs, pregnant mice were sacrificed at day 13-13.5 of gestation, fetuses were dissected and the heads, livers, mesonephros, and hearts were removed. The remaining parts, including limb buds and body wall were minced with a scalpel blade, incubated in trypsin/EDTA solution for 20 min at 37°C and disaggregated to single cells and clumps by vigorous pipetting after adding warm culture medium. The culture medium used was high glucose DMEM (Lonza), supplemented with 15% fetal bovine serum (FBS) (Gibco, Lot 41Q2035K), non-essential amino acids (GE Healthcare), sodium pyruvate, 2 mM Glutamine (GE Healthcare), and 0.1 mM 2-mercaptoethanol. Feeder cells were produced by treating MEFs from NMRI mice with 10 µg/ml mitomycin C for 2 hours.
Reprogramming expression vectors
The construction of the Sleeping Beauty (SB) transposon plasmids with either the Tet-On (TetO) promoter (SB-TetO-pOSMK-IRES-Tomato, used together with SB-CMV-rTA-IRES-Neor) or with the constitutive CAG promoter (SB-CAG-pOSMK-IRES-Tomato) for expression of the porcine OCT4, SOX2, c-MYC, and KLF4 have been described previously[16]. To create a CRE-excisable version, the CAG promoter and the polyadenylation signal were amplified using loxP-containing forward or reverse primer, respectively, and these modified sequences were used to replace the originals by standard DNA cloning techniques, thus generating a loxP-flanked CAG-pOSMK-IRES-Tomato-pA cassette. For efficient integration into MEFs, the plasmid with a hyperactive version of the SB (SB100x) was co-transfected with each of the reprogramming transposons. The SB transposon vector backbone and transposase expression plasmids were provided by Dr. Zoltan Ivics (Paul-Ehrlich-Institute, Langen, Germany).
Mouse iPSC generation and culture
Three different MEF lines from C57BL/6-OG2 mice were electroporated at passages 3-4 with the SB transposon and transposase plasmids as described by Petkov et al. [16] and plated on gelatinized T75 flasks. The cells were cultured in MEFs culture medium for 2-3 days and then were then split 1:3-1:4 to T75 flasks with mitotically inactivated MEF feeders and the medium was supplemented with 1000 U/ml ESGRO murine leukemia inhibitory factor (mLIF) (Millipore). When vectors with the TetO promoter were used, the medium was supplemented with 5 µg/ml DOX. The OCT4-EGFP-positive colonies with ESC-like morphology were manually picked at day 10-11 post-transfection, disaggregated with Trypsin/EDTA, and cultured individually on mytomycin C-inactivated MEFs to establish individual cell lines. The established miPSC were regularly split every 3-4 days at ratio 1:15-1:20 by trypsinization and maintained on fresh feeders in ESGRO medium for over 70 passages. For generation of transgene-free miPSC, the miPSC harboring the “floxed” version of the reprogramming transposon were transfected with SB-CAG-CRE-IRES-Puror vector using Lipofectamine 2000 (Life Sciences) as instructed by the manufacturer and selected with culture medium supplemented with 3 µg/ml puromycin.
Alkaline phosphatase (AP) staining and immunocytochemistry
The cultured were fixed with 4% Formalin solution for 5 min (for AP) or 15 min (antibody staining) and washed with PBS prior to further processing. For AP staining, a solution containing 1 mg/ml Fast Red and 0.4 mg/ml Naphtol-BI-Phosphate in AP-buffer (100 mM NaCl2, 100 mM Tris (pH 9.5), and 50 mM MgCl2) was applied to the fixed wells. The AP-positive cells were discerned by the formation of insoluble red-brown precipitate. The presence of SSEA-1, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81 expression in miPSC was assessed by incubation with primary antibodies (Thermo; Cat. numbers 41-1200, MA1-020X, 41-4000, 41-1000, 41-1100, respectively), diluted in PBS with 2% Knockout Serum Replacement (KSR) (Invitrogen) at concentration 10 ng/ml for 1 hour, followed by washing with PBS and incubation with the secondary peroxidase - conjugated antibody. For staining of neuronal-like cells, the anti-PGP 9.5 primary antibody (DACO) was used in a similar protocol. The expressed proteins were visualized after incubation with peroxidase substrate from AEC staining kit.
Reverse transcription, PCR, and real-time PCR For RNA extraction, the miPSC cultures were washed 3 times with Ca- and Mg-free PBS and incubated in the same solution for 15 minutes at 37 degrees C. The miPSC colonies were detached from the feeders by gentle pipetting, collected in a 15 ml centrifuge tube and allowed to settle on the bottom of the tube for 10 min. The supernatant containing single cells (presumably mostly feeder MEFs) was carefully removed, the pellet was re-suspended in PBS and the colonies were pelleted by centrifugation, lysed in TRI reagent (Ambion), and total RNA was extracted as instructed by the manufacturer. Reverse-transcription was carried out with MuLV Reverse Transcriptase (Applied Biosystems). Polymerase chain reaction was performed using 50 ng reverse transcribed RNA per 25 µl reaction volume using Platinum Taq Polymerase (Invitrogen).
Real-time relative quantitative analysis was run on the ABI 7500 Fast System using SYBR Green Master Mix (Applied Biosystems). The primers used for PCR and real-time PCR analysis of endogenous mouse pluripotency markers Oct4, Sox2, Nanog, c-myc, Klf4, Rex1, and Utf1 as well as for the differentiation markers alpha fetoprotein (Afp), Sox17, Gata4, Gata6, cardiac troponin, myosin heavy chain (MHC), nestin, β-III tubulin, and Pax6 are shown in Table S1. The primers used for the amplification of the mouse endogenous pluripotency genes were specific only for the mouse cDNA sequences and did not amplify the pig pOSMK cassette (results not shown). The primers used for the amplification of the pOSMK-IRES-Tomato construct were a forward primer binding at the end of KLF4 and a reverse primer binding to IRES (shown in Table S1). The data from the relative quantitative analysis were analyzed with GeneEx software (bioMCC). Statistically significant differences were determined by Student’s t-test performed by the same computer program.
Karyotyping
Three lines each from TetO- and CAG miPSC at passages 58-60 were treated with Demecolcine solution (Sigma, D1925) added at 100 µl/10ml culture medium for 35 min. The cells were harvested by trypsinization, incubated in hypotonic solution (0.28% KCl + 0.25% Na Citrate) for 25 min at 37˚C, and fixed in cold (-20˚C) methanol: acetic acid (3:1) fixative for 1 hour. Metaphase spreads were produced by dropping 50 µl drops onto glass slides placed at a slight angle over steaming water bath (95˚C) and waiting 1 min before removal and air drying. The metaphase spreads were stained with 2% Giemsa solution. The metaphase chromosomes of at least 15 metaphase spreads per cell line were counted to determine whether the cells had normal karyotype.
Embryoid body (EB) formation
Mouse iPSC colonies were separated from the feeder as described above in subsection 2.5, disaggregated to single cells with trypsin, and suspended in basic culture medium without ESGRO. “Hanging drops” were then produced by making 25 µl drops on the inner side of a 10 mm Petri dish lid (containing 2000 cells/drop) and placing the lid back on the dish filled with PBS (to prevent drying of the drops). After 5 days in “hanging drops” culture, the emerging embryoid bodies were plated in gelatin-treated 6-well plates and cultured further for 15 days with changes of the medium every 3-4 days. For neuronal differentiation, the attached EBs were cultured in N2 medium (DMEM/F12 supplemented with sodium pyruvate, amino acids, penicillin-streptomycin, and N2 supplement (Thermo)).
Teratoma formation
Prior to conducting the animal experiments, permission was obtained from the local animal welfare authority (LAVES).
Nude immunodeficient mice were produced by mating outbred NMRI strain parents. A total of 13 nude mice (7 mice/TetO group and 6 mice/CAG group) were injected each with 1 x106 putative iPSC derived from reprogramming with the porcine vectors subcutaneously in the right flank. The mice were observed daily for 3 weeks. After reaching the critical size of the tumour (1 cm), the animals were sacrificed. Biopsies were taken for DNA/RNA extraction, and tissues were fixed in neutral buffered 4% formalin not exceeding 48 h, dehydrated (Shandon Hypercenter, XP) and subsequently embedded in paraffin (TES, Medite). Sections (2-3 μm thick, microtome Reichert-Jung 2030), were deparaffinized in xylene and H&E stained according to standard protocols. The morphological evaluation for the presence of the three germ layers (Axioskop 40, Zeiss microscope) was performed by a trained pathologist and representative microphotographs were taken (AxioCam MRc, Zeiss).
Results
Reprogramming to pluripotency Transfected MEFs cultured on inactivated feeders formed O4-EGFP-positive colonies with typical miPSC-like morphology within 7-8 days post-transfection (Figure 1A, D). A number of granular colonies consisting of round, highly refractory cells were also observed.
Figure 1 –Mouse iPSC generated with Sleeping Beauty transposons carrying porcine reprogramming factors. A) TetO-miPSC at day 8 post-transfection with typical miPSC morphology. B) Oct4-EGFP reporter expression in the colony in image A. C) Silenced pOSMK-IRES-Tomato expression except in some cells in image A. D) CAG-miPSC at day 8 post-transfection. The colony has compact miPSC-like morphology. E) Expression of Oct4-EGFP reporter in the colony shown in image D. F) Expression of non-silenced pOSMK-IRES-Tomato construct in the colony from image D. G) CAG-miPSC at P.22. The colonies maintained compact miPSC morphology. H) Oct4-EGFP reporter expression in the colonies shown in image G. I) Expression of pOSMK-IRES-Tomato in the colonies shown in image G. (All scale bars = 50 µm).
There was no significant difference in the timing of EGFP-positive colony appearance between the different groups where transposons with TetO or CAG were used. In most miPSC colonies produced with the

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Additionally, this is a varied label listing. This means the coins could come with additional labels in the slab such as Early Release or UCAM. However, this is not guaranteed. All coins are chosen at random when your payment is processed. If you purchase more than one, please keep in mind you could receive several coins that have the same additional labels, varying labels, or no labels at all.
Walking Liberty graces the obverse of 2019-S Silver American Eagle Enhanced Reverse Proofs. This iconic design was created in 1916 by Adolph A. Weinman and featured on the US half dollar until 1947. It captures the spirit of hope for the future that existed in the early 20th century as Liberty walks toward the sun on the horizon extended her hand to the warmth and light of a new day.
The heraldic eagle of the United States features on the reverse of 2019 1 oz Silver American Eagle Enhanced Reverse Proofs. This emblem was first created in 1792 by Robert Scot and modified for this series in 1986 by John Mercanti.American Eagle 2019 One Ounce Silver Enhanced Reverse Proof Coin Mintage
These 2019-S 1 oz Silver American Eagle Enhanced Reverse Proofs, generally speaking, have frosted fields and mirrored designs. As an enhanced reverse proof, the US Mint selectively applied frosted and polished fields to isolated design elements for a unique visual appearance. Each coin bears the “S” mint mark of the San Francisco Mint on its reverse field.2019 American Eagle One Oz. Silver Enhanced Reverse Proof Coin
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Free Online Blackjack: Play for Free or Real Money in 2021 Online blackjack is one of the most popular casino games in the world. Not only is it simple to play, but the best online blackjack games offer great odds that few other options can match. The exciting game of Blackjack is the world’s most popular casino game, and now you can play it for real money prizes online at CaesarsCasino.com! Also called ’21’, Blackjack online comes in many variations, but the rules are all very similar and easy to follow.
Blackjack is arguably one of the most popular casino games. You can find online blackjack, daily blackjack tournaments at casinos, multi-player blackjack, and even Facebook blackjack. Below we recommend some of our top casino picks for real money blackjack. For those of you interested in money limits, we also recommend the best casinos for high limit blackjack games as well as low limit $1 games.
And while it’s not real money your free blackjack online winnings will earn you treats that you can use for fun stuff. Where else can you go to play free blackjack online and get something in return? So take a moment, place your bets and get ready to play our free blackjack game. Great Day Games + Free Games = Lots of Fun. Free blackjack casino game is the perfect way to practice your blackjack strategy before hitting the real money table. Play blackjack online, free of charge on this page and find your favorite virtual blackjack for fun. Online blackjack free play modes are aimed only at adults, and you must over 18 years old to play blackjack games even for fun. Free blackjack games are played with virtual chips without any real-money value. You play with the free chips you get when you first load the game, and these can’t be exchanged for real money, regardless of how many chips you accumulate during your free play.
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Sweepstakes casinos use digital coins instead of real money as currency. Players accumulate digital coins by winning games and can later exchange them for cash prizes. Because sweepstakes casinos are not legally classified as gambling, they are popular among players who live in states that have not yet legalized real money online gambling. The sweepstakes format provides a similar, but legal alternative to the online casino experience.
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Chumba Casino is a great option as it offers both slots and table games. Its variety is what makes it a unique sweepstakes casino.Free Blackjack Online
Blackjack is a fun game to play, whether you have real money on the line or not. While the idea of playing for no money may appear pointless to some hardened gamblers, it is essentially no different to someone playing Angry Birds or watching a movie, you are simply using your time to entertain yourself.
Apart from entertainment free table games are useful for practicing for real money games and weeding out mistakes as I have covered here. I am also letting you know of the benefits that can be found by using a blackjack training programs. These are also free games with the added advantage of letting you know what you are doing wrong. After that, I have to mention some minor downsides to playing free blackjack – before summarizing some of the best places to find free blackjack games for a number of devices right at the end.How Free Blackjack Can Develop Your Real Money Games
The main benefit you can gain from playing free blackjack is getting used to the game itself. You can get a feel for how the software of a casino site operates by trying it out before you play for real money. If, for example, you were playing for real money and incorrectly raised your bet size without realizing, it could cost you potentially a lot of money. By playing for free money you can make these mistakes for free then move over to real money games when you are used to everything.
Free blackjack games also give you the opportunity to perfect your skills. While basic strategy can easily be followed when playing real money you may feel pressured, especially if you are playing live. By giving yourself chance to practice in play money, you are learning you base skills without risk and added pressures. Card counting should also be tested on sites with suitable games, such as those with few or even single decks, as you do not want to be looking obvious in a casino.
If you are developing new betting strategies, it makes sense to test run them first. Again the no risk nature of the free blackjack games gives you the opportunity to try new things without consequence. An additional bonus when testing new ideas is that you can get through hands far quicker online than if you were dealing them out physically, giving you a much bigger sample size to base results on.
Finally by allowing you to play for free, many casino sites, such as Spin City Casino and Casino Titan give you the chance to try out the different type of blackjack variants they host.Free Games to Learn With
For the majority of experimentation and entertainment a simple free game will suffice. If you wish to get instant feedback on how you are playing you can find blackjack trainers online. You can purchase these from retailers or find simple custom designed programs online which individuals have made for their webpage or blogs. These are normally built to a simple design in order to explore the math and strategy behind blackjack.
A basic strategy trainer plays like a free game and alert you when you break away from basic strategy before letting you know what the correct play was. Some do this with an alarm, while others display percentages of wins or loses. Some may include both. If you are new to the game having a trainer program for basic strategy will shorten the learning curve considerably.
Card counting trainers also exist. While practicing card counting with real cards is preferable, online trainers have the advantage of being convenient to play hassle free at any time and a computer’s ability to hold a count without making mistakes.Free Blackjack – The Negatives
While some may find blackjack an entertaining game by itself, the game is made far more interesting when it is played for money. The added risk and reward of actual money does add another dimension of excitement to blackjack, it is after all designed as a casino game. Without the risk of winning or losing anything to hold attention, rather than playing blackjack the game can easily become a thoughtless exercise of losing time and play chips. Small limit blackjack sites like William Hill offer games as low as 10p, so there are games running that everyone can afford and enjoy.
By playing for free money you are not eligible for the rewards real money can bring, apart from the winnings. All online sites offer bonuses for real money deposits which is free money you unlock by playing on the site. If you play live you can benefit from comps and rewards in your casino, entitling you to anything from free drinks to reduced room rates or even more if you are a high roller. This way you can get more for your time that you will not get playing free games.Keeping It Fun, Where to Find the Best Free Blackjack Games
There are plenty of free blackjack games available online. Most are often online casinos giving you a taste of what they offer before you go ahead and deposit, others may be training tools to help you improve your game.
Nowadays, there is a whole new genre of free blackjack games which you can play socially – either with your friends or with new people. These games come in app format, or via the many Facebook games, and often include challenges, leader boards or gifts inside the games. The app creators would like you to buy more chips, though most top you up daily without needing to pay.
Regardless of how you use these free games, even if it is a fun time-waster, I hope you continue to enjoy the game of blackjack.Best Online Blackjack For Real Money
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The purpose of this page is to serve as an introduction to the game of blackjack. It includes information about the history of the game, how to play, and how to win. Blackjack is one of the most popular casino games in history, and this is due in no small part to the element of strategy involved. Players in blackjack make decisions, and those decisions affect the outcome of the game. Most other casino games don’t offer this level of control.The History Of Blackjack
Most people don’t think of Cervantes’ novel Don Quixote when they think of blackjack, but it’s possibly the first mention of the game in literature. That’s quite a distinction when you consider that Don Quixote is also the first novel ever written—prior to Don Quixote, poems and plays were the primary literary form throughout the world.
The game mentioned in that book wasn’t called blackjack, though—it was called “ventiuna.” Little is mentioned about how the game was played in the book, but enough is discussed to make it clearly recognizable. The goal of the game, as described in the book, is to get a total of 21. Also, the book makes it clear that aces could count as either 1 or 11. It’s hard to find a more recognizable description of blackjack than that.
The game of blackjack grew popular because it spread via various port cities throughout the world. The name changed with the country, and so did some of the details of gameplay, but a rose by any other name would still smell as sweet. For example, the British called the game “pontoon.” The game spread to the United States in the 1800s, and it traveled west with the rest of the country.How To Play Blackjack
Blackjack is an easy enough game to play. My mom taught me the basics when I was ten years old.
The player and the dealer are each dealt two cards. All cards have a point value equal to their ranking. Face cards (the jack, queen, and king) are each worth ten points. An ace can be counted as 1 or 11, depending on which would be more favorable to the player.
The goal is to get as close as you can to 21 without going over. If your total exceeds 21, you lose immediately.
The main decision you make in a game of blackjack is whether or not to take another card. Taking a card is called being “hit,” while not taking another card is called “standing” or “staying.”
Rules variations at casino offer a couple of extra decisions to players. For example, players are often allowed to “double down,” which is a variation of hitting in which you take one (and only one) more card while doubling the size of your initial bet. Players are also allowed to split pairs by placing an additional bet. (A pair consists of two cards of the same rank—two kings, for example.) This creates two one-card hands which immediately are dealt an extra card to form two new hands of blackjack.
Most casinos in major gambling destinations like Las Vegas, Reno, and Atlantic City offer “classes” on how to play blackjack during the slower-traffic periods on the gaming floor. Taking one of these classes in order to learn how the game plays is an excellent way to get a feel for the game.
Another great way to learn how to play blackjack is to explore some of the free blackjack games that are available online. You can play for fake chips in order to understand how the action works, who goes first, and what order everything happens in.How To Win At Blackjack
“Winning” means different things to different people. Some people feel like they’ve won even if they’ve lost a couple hundred dollars. Others just want to get lucky and put some money in their pocket. In any gambling game, there’s no sure way to book a win every time you play. These are games of chance, after all. But there are ways to win more often and lose less money over time.
The first step to becoming a winning blackjack player is to study the game and learn how to play. For example, a common novice mistake is to buy in at a table where the minimum bet is larger than their bankroll can afford. If you buy in for $100 at a $50 minimum table, you can only afford to lose twice in a row. Some people are so embarrassed by this mistake that they just go ahead and lose.
If you don’t understand the game well enough to keep up with the totals, you’re going to lose a lot more money than someone who knows the rules and gameplay inside out. This is one of the reasons I suggested playing some of the free games online.
The second step to becoming a winning blackjack player is to learn basic strategy. This step alone will reduce the house’s edge from 4% to less than 1%. (The house edge is the amount that the house expects to keep over the long run because of the rules of the game.) Basic strategy is the mathematically correct decision in every situation that comes up at the blackjack table.
Most people encourage players to memorize basic strategy using a chart or a table of some kind, but my preference is to learn the rules for how to play each hand individually. David Sklansky’s book, Sklansky on Blackjack, takes this approach, describing how to play every possible hand in every possible situation by looking at each possible total you could have.
Once you’ve reduced the house edge to less than 1% by using perfect basic strategy, you can start to take advantage of the many comps that casinos offer players. You can receive food, event tickets, and even travel perks just by making sure the casino tracks your play. These rewards have a monetary value, so even though you have a negative expectation against the casino, you get something back, making that negative expectation ever smaller.
The third step to becoming a winning blackjack player is to start learning some advantage gambling techniques like card counting. Once you’ve mastered a card counting system, you can actually get an advantage over the house. Counting cards isn’t as hard as you might think, either. Anyone who’s willing to put in the time and effort can learn a simple count like the Hi Lo Count or the Ace Five count.
Blackjack is a great game. It’s fun, it’s a mental challenge, and it offers some of the best math in the casino. If you like making decisions that affect your outcome, blackjack might be the perfect game for you.Casinos With Daily Blackjack Tournaments
Blackjack tournaments are a new twist on an old casino gaming favorite. The basic rules stay the same – you are trying to beat the dealer’s hand without going over 21. What is different is that you are playing against other gamblers and not the house. Your objective in a blackjack tournament is to accumulate more chips than the other players at your table – and move on to the next round. This article covers the setup and strategy for blackjack tournaments, which you can now enjoy online as well as in live casinos.
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*Choice of First, Club World, World Traveller Plus and World Traveller. (First available on some aircraft)Photo galleryTechnical information Boeing 777-200 Number in fleet45Passenger capacityup to 336 (3 or 4 class)Length63.7m (209 feet 1 inches)Wingspan60.9m (199 feet 11 inches)Height18.5m (60 feet 9 inches)Engines2× Rolls Royce Trent 800 or 2× General Electric 90Maximum speed950km/h (590mph, Mach 0.89)Range6,857km (4,261 miles)Seating plans
These are the different seating layouts for the Boeing 777-200:

*4 class aircraft (with a First cabin)
*3 class aircraft (without a First cabin)
*3 class aircraft (without a First cabin and available on selected LGW routes)Did you know?
*The diameter of General Electric engines on the Boeing 777 is as wide as the passenger cabin of the Boeing 737.
*In 2006, British Airways set a new record for the longest non-stop commercial flight. The Boeing 777-200 flew 17,157km (9,274nm) from Brussels to Melbourne, in 18h45.Improvements for London Gatwick fleet
If you’re flying from London Gatwick, you can now look forward to a fully refreshed Boeing 777 fleet and new generation economy (World Traveller) and premium economy (World Traveller Plus) seats. The refreshed cabins will soon start appearing on the Boeing 777 fleet at London Gatwick and the whole fleet will be updated by Summer 2019.
The economy and premium economy cabins have been fitted with elegant new seats with 50% larger entertainment screens and each seat will have a USB socket. Work from home bingo.777x Aircraft777er Vs 777lrWhere does the Boeing 777-200 fly?777.beThe aircraft operates on the following routes:
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Our guide is aimed to explain UK-based players how to play legally in online casinos even if they’re registered with Gamstop. You’ll also get a few brief reviews of the best gambling sites.
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*Under the newly updated gambling laws in the United Kingdom, any casinos not on GamStop and online poker providers should not be allowed to offer services. The good news is that there are exclusive UK poker sites not on GamStop self-exclusion such as BetOnline, Fortune Clock, Playhub, and other poker rooms.
*Welcome to casinosnotongamstop.uk, a portal dedicated to finding the best and most trustworthy casinos and Gamstop gambling sites not registered with Gamstop.Here you will find slot sites, bingo sites, betting sites and all round non gamstop casinos. We have carefully vetted all gambling sites that are listed on our website, and would consider all listed here as trustworthy gambling sites.Content
Instead, you will come across a lot of non-GamStop betting sites that are licensed by other respected authorities, such as the Malta Gaming Authority. Other common regulators include Gibraltar, Alderney, and Curacao. Bookies Bonuses. A lot of the time, you will come across bigger and better offers when you are using non-GamStop sites. This is because there are fewer. As a new online casino not on Gamstop, Harry’s Casino is one of live non Gamstop casino sites and has a lot of table games to choose from. Players aren’t just limited to the traditional American and European versions of blackjack and roulette. From 3d computer-generated graphics to live dealer games, Harry’s Casino has it all.The Best Casinos Not on Gamstop 202195.0%Fortune Clock Casino50 FREE SPINS on Starburst100% up to €1,000 and 100 extra spins (€0.1/spin)75% up to €1,000 and 75 extra spins (€0.1/spin)50% up to €1,000 and 75 extra spins (€0.1/spin)18+ ∙play responsibly ∙To the casinoRead the review 92.0%Jackpot Charm CasinoBonus 150%Bonus 125%Bonus 100%Bonus 75%Bonus 50%18+ ∙play responsibly ∙To the casinoRead the review 90.0%Agent NoWager Casino5 EUR for Sign Up10% up to €1,000 (Wager: 1x bonus)12% up to €1,000 (Wager: 1x bonus)15% up to €1,000 (Wager: 1x bonus)18+ ∙play responsibly ∙To the casinoRead the review 89.0%Magic Reels Casino200% up to 150 EUR50% up to 300 EUR75% up to 300 EUR18+ ∙play responsibly ∙To the casinoRead the review 87.0%BetOnline100% up to $1,000100% up to $1,000100% up to $1,00018+ ∙play responsibly ∙To the casinoRead the review 86.0%WG Casino5 EUR200% Bonus100% Bonus150% Bonus18+ ∙play responsibly ∙To the casinoRead the review 85.0%Triumph Casino100% Bonus125% Bonus150% Bonus18+ ∙play responsibly ∙To the casinoRead the review 85.0%Very Well Casino25 free spins in Book of Ra75 free spins in Dolphin’s Pearl100 free spins in Lord Of The Ocean125 free spins in The Money Game50 free spins in Lucky Lady’s Charm18+ ∙play responsibly ∙To the casinoRead the review 78.0%BetNow Casino50% up to $1,00050% up to $1,00050% up to $1,00018+ ∙play responsibly ∙To the casinoRead the review 63.0%Casino Intense10€ Bonus100% up to €10050% up to €100 Weekend Reload Bonus100% up to €200, 3rd Deposit Bonus50% up to €588, 2nd Deposit Bonus18+ ∙play responsibly ∙To the casinoRead the review 55.0%Wild Vegas$60 for Free350% up to $3,500300% up to $90 and 30 extra spins ($0.1/spin)18+ ∙play responsibly ∙To the casinoRead the review 53.0%Crazyno Casino100% up to €500 and 50 extra spins (€0.1/spin)50% up to €500 and 50 extra spins (€0.1/spin)100% up to €500 and 50 extra spins (€0.1/spin)18+ ∙play responsibly ∙To the casinoRead the review 52.0%Crazy Star Casino150% up to €765175% up to €765200% up to €76550% up to €76575% up to €765100% up to €76518+ ∙play responsibly ∙To the casinoRead the review 46.0%Ph CasinoBonus 100% up to £500 (Promo Code: FIRST)18+ ∙play responsibly ∙To the casinoRead the review
Gambling is usually fun and entertaining, and it can add hundreds and thousands of pounds to your wallet if you’re lucky. Still, it can also be very addictive for some people and become a serious problem. Such gamblers need help and support, and that’s where Gamstop can be useful for them.
Gamstop is a self-exclusion scheme that gives aid to people with addiction to online casinos. This program helps them to give up gambling by blocking their access to all the UK casinos that are licensed by the UK Gambling Commission. Since you’re registered with Gamestop, you won’t be able to play on any of these websites using your existing accounts, as well as creating a new one.
The program is really helpful for many people, but some gamblers registered for it by mistake and they regret their decision. Some of them wanted just to get rid of one particular gambling site, others just didn’t realize all the consequences of joining the self-exclusion scheme.How to play online casino games if you’re registered with Gamstop
If you’ve signed up for the program by mistake, you can still gamble online. The best way to do that is to join non Gamstop casinos. Such websites don’t follow the regulations of this self-exclusion scheme, so you can sign up freely and play all your favourite games.
Note: You shouldn’t try to get rid of Gamstop this way, in case you feel even the slightest addiction to casino games or if you have a history of problem gambling. The service was launched to help people and it’s not wise to reject the aid it provides.
Still, if you feel that gambling has no bad influence on you and you can control how much you spend on games and betting, then casinos not on Gamstop are a good choice for you. They not only allow you to play again after you’ve self-excluded yourself but also give you lots of benefits like generous bonuses and high limits on withdrawals and deposits.
If you’re in search of the best non Gamstop casinos, you can use the rating below. It gathered all the top gambling sites not using Gamstop but accepting UK players. We’ve rated these casinos according to the number of games they provide, bonuses they offer, deposit/withdrawal policy and users feedback. You can pick any site in the list and be sure that it’s safe to play and you’ll get incredible gambling experience.
Once you’ve signed up for one of these international casinos not registered with Gamstop, you’ll be able to play freely all the games available, including slots, blackjack, poker, roulette, bingo, craps, baccarat and other games not on Gamstop. Moreover, you’ll get a lucrative bonus reward for the first deposit on the site, as well as for a few following top-ups. Lucky players that we’re fortunate to win big prizes will be able to obtain all the money fast due to high limits on withdrawals.What is Gamstop and how does it work?
First of all, Gamstop is a great web service that helps gamblers to fight their addiction to casino games. It offers players support via live chat or phone, as well as the ability to register on their website and block yourself from all the gambling sites in the jurisdiction of UK Gambling Commission.
The program is operated by National Online Self-Exclusion Scheme Limited, which is a non-profit organization that was founded in 2016. “NOSES LTD.” company launched its website in 2018 to allow all the gamblers that have issues with betting and casino games to get rid of their addiction. Gamstop is free for players and everyone can join it in about 10 minutes.
The service gives you 3 options to block yourself from gambling sites:
*half a year;
*one year;
*five years.
The first options are suitable if you want just to have a break and then start playing casino games again. They’re good to restore the control over the time you spend on gambling and the amount of money you bet. The last option is preferable for people who are strongly addicted to betting and online casino games. If you choose it, then you’re aiming to break up with online gambling forever. Choose wisely depending on the goals you’ve set for yourself.How does Gamstop work?
The program exploits a simple, but a very efficient method to prevent gamblers from using gambling websites. It collects your personal data when you sign up and then casinos-participants use this information to block the appropriate users’ accounts and disallow them to create new ones. Since January 2020 Gamstop is mandatory for all the UK casinos. They should join the program if they want to get a new license or not to lose the old one.
When the player registers with Gamstop, he/she can be 100% sure the access to British online casinos is impossible. No website will disobey program guidelines, cause the penalties are as strong as they can be. It can be a huge fine, which is the slightest punishment, or the casino can just lose its license. No casino operator will risk its business to give blocked players access to gambling sites.
If you’ve decided to sign up for Gamstop, the first step is to visit their website. You’ll have to register online and provide such personal data as the date of birth, email and postcode. The whole process will take about 10 minutes. All you need is just to follow the instructions.
The schemes start to work as soon as you’ve signed up. Still, it usually takes up to 24 hours before all the participants will get new information and your access to gambling sites will be blocked.
All the terms and conditions are explained on the Gamstop website. Still, some things may be unclear. Here are the answers to some common questions:UK Non Gamstop Bookies > Best Betting Sites Not On Gamstop
*I have money on my account, how can I get them back? There are a few ways to do that. The simplest option seems to be the email to the casino support. Explain the issue and they’ll definitely cooperate. The other method to get your funds back is just to call them and ask how you can return the money. At last, you can just visit the offline casino that operates the site (if any) and ask for the aid with your issue.
*Is it possible to reverse Gamstop? Can you undo Gamstop? The short answer is Yes, but it’s very complicated and the chances for success are rather low. To do that you’ll have to contact the program’s team and persuade them that you’ve joined the self-exclusion scheme by mistake. Don’t even try to fool them, cause these people are leading specialists in their sphere.
*Does Gamstop affect credit rating? Neither the service itself nor any casino-participant has access to your bank account or the ability to influence on your credit rating. The only thing that really affects it is the fact that you’re using borrowed funds to play casino games.
*How to cancel Gamstop? There is no way you can cancel Gamstop. You should just wait until your self-exclusion period is over and then contact the service and ask them to delete your information from their database.
*Are there any alternatives? Yes, there are lots of services that offer you the tools to protect yourself from gambling addictions, including GamCare, Gamban, Netnanny etc. There are many casinos you can play at although you are on Gamstop.
*Is Gamstop free? Yes, you won’t have to pay for anything.
If you think gambling has a bad influence on your mental health and it’s addictive, then Gamstop is probably the best online service to help you with your issue.How to get around Gamstop?
There are a few methods that will potentially work. Some players create new accounts using their spouse’s personal data. Others try to sign up by altering contact information and name. You can also join online casinos without users verification.
Still, you definitely have problems with online gambling if you’re going to try one of the methods mentioned above. There is another way to play all the casino games you want without trying to fool someone or hide your identity. Just join sites not blocked by Gamstop. They’ll allow you to gamble again and offer lucrative bonuses for new players.
Note: Casinos not on Gamban are licensed outside the UK, so you can access them freely even during your self-exclusion period. However, you shouldn’t do that if you have any problems with gambling or you feel that you have no control over how much you play and bet. Remember that you’ve registered with Gamstop for a reason and it’s a very bad idea to neglect all the help and support you’ve received.Types of gambling sites not using Gamstop
The sites not covered by Gamstop are licensed off-shore and they accept players from the United Kingdom. Here are the major types of such websites:
*Casinos sites not on Gamban. It’s the most popular type of gambling sites not linked to Gamstop. They offer their users lots of games in different categories, including slot machines, table and card games, scratch cards, bingo and live games. New players can get lucrative welcome bonuses that will match their deposits up to 500% in total and free spins for slot games. Casinos that don’t use Gamstop are licensed by off-shore authorities like MGA or Curacao E-gaming.
*Poker sites not on Gamstop. If you like such games as Texas Holdem, Omaha and 7-Card Stud, then you should definitely sign up to one of poker gambling sites without Gamstop. You’ll be able to play all your favourite card games again and participate in tournaments. Such poker rooms will also offer generous free chips bonus for new players.
*Bingo and lottery sites not on Gamstop. Bingo is one of the most popular games in Great Britain and its popularity seems to increase year after year. It engages more and more players due to its simple rules, big prizes and “the magic of numbers”. It’s more than exciting to shout “Bingo” when you have all the 5 numbers in a row and get a win. Bingo sites not on Gamstop will give you the opportunity to get “Bingo” again.
*Slots not on Gamstop. Gambling sites that are focused on slot machines will offer you hundreds of games to choose from, including fruity slots, classic reels, video slots and Jackpot slots. This game type is popular among players for rich payouts, special features, bonus rounds and rather high RTP level. Slot machines give you a chance to win tens of thousands times more than your initial bet.
Slots gambling sites will offer you generous welcome bonuses and lots of free spins if you’ll register and start playing online.
*Lotteries. All the lotteries are the games of pure chance. You don’t need to apply any strategies and learn complicated rules. Just make your bet and wait for the outcome. If you’ll join gambling sites not registered with Gamstop, you’ll definitely find all your favourite lotteries there.
*Sports betting sites, not on Gamstop. Bookmakers that are not licensed in the UK will give you lots of betting options to choose from. Bookies not on GamStop have all the major sports available and you’ll be able to wager on the outcome of matches in national leagues, world cups, European tournaments etc. Be aware that these bookmakers are not focused on players from the United Kingdom, so a few popular in GB tournaments may be missing at these sportsbooks.
You can be 100% sure that websites without Gamstop have all the games you can play in UK casinos that are licensed by UK Gambling Commission. If you need a particular slot or table game, just check whether it is available on the website before you’ll register. Casinos not on Gamstop also offer a wide range of live games where you play with the real dealer online.How to choose the best non-UK casino sites?
There are lots of great gambling sites without Gamstop, but there are also the ones you should stay away from. If you want to pick a decent casino without Gamstop, make sure it meets all the requirements listed below:
*the website you sign up for has to be licensed by one of the major industry authorities. It can be Curacao E-gaming, Malta Gaming Authority, Kahnawake Gambling Commission etc. Also, make sure the casino has implemented all the responsible gaming rules and uses Random Number Generation to provide fair results in games;
*top non Gamstop casinos offer their users a big variety of games in different categories, including live games. The software is usually provided by such famous developers as NetEnt, Evolution, Play’n GO, BetSoft etc;
*the best casinos not on Gamstop will provide lucrative 1st — 4th deposit bonus to new players. You’ll be able to receive prizes that are up to 500% bigger than the top-up amount. Moreover, you can get free spins for slot machines and chips for table games;
*the website you’re going to join has to give you the opportunity to deposit and withdraw funds fast and secure. Also, make sure the limits are high enough so that you can transfer to/from your account the amount of money you want;
*non Gamestop casinos are based outside the United Kingdom and their main currency is usually Euro or US dollar. Still, there are the ones that allow you to open an account in GBP. Join such gambling sites not to pay commissions for currency exchange.
You can play in casinos, not on Gamstop and win really big prizes, but you should be careful with the fact that your rewards may be taxable if the website is licensed outside the European Union. That’s a good reason to search for casinos not registered with Gamestop that have licenses from EU authorities.What are the main benefits of non Gamstop casinos?
Well, the first and the most important one is that they allow you to play online even if you have self-excluded yourself using Gamstop program. You can access all the games you like, make bets and win big prizes without any limits.
Still, casinos not under Gamstop have way more benefits. Since they’re not licensed in the UK and they don’t follow the guidelines of the program, these websites can offer their users way better bonuses and lower wagering requirements. Casinos increase prizes to obtain more customers, while you can benefit from it by getting lots of free credits and spins to play online for real money. Moreover, you’ll play through the prizes very fast due to low wagering requirements.
Non-Gamstop casinos are free from UK regulations, so they can offer you various gamification features and way higher limits on depositing and withdrawing funds.
You can easily join one of the casino sites not affected by Gamstop just in a few minutes. To do that, visit the official website, click the signup and enter all the appropriate data in the registration form. Then you’ll have to make a deposit to get bonuses and to play for real money. There are lots of payment systems available for that purpose, including debit/credit cards, e-wallets, Bitcoin, prepaid cards and Bank transfer. You may n

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Jul 24, 2017 I thought it might have been acid damage as well so I took quarters of the same year-and subjected them to 3 different kinds of acid hydro chloric sulfuric and nitric None of the coins resulted in pitting Only disolving the oxidized layer. The 1965 Doubled Die Obverse Quarter Dollar (FS-101) is a very rare and popular variety that shows clear doubling on most of the letters of IN GOD WE TRUST, LIBERTY, the date and on the left side of Washington’s profile. Clad quarters weigh 5.67 grams. The present coin, which is made of 90% silver, weighs 6.26 grams, the approximate weight of the 1964 quarter. Evidently a sliver planchet remained in a tote bin crevice and later was dislodged and mixed in with the clad planchets. This is the first year in which the Mint struck Clad Washington Quarters. The 1965 circulation strike Quarter is really common in most grades. It is considered somewhat scarce in MS65 condition. In MS66 it is scarcer with a few thousands examples probably in existence. The quarter is a Canadian coin, valued at 25 cents or one-fourth of a Canadian dollar. It is a small, circular coin of silver colour. According to the Royal Canadian Mint, the official name for the coin is the 25-cent piece, but in practice it is simply called a quarter.
*1965 Quarter Value Silver
*1965 Quarter Value Worth
*1965 Quarter Value Liberty > > > > >
NOTE: Many features on this website require Javascript. You can enable both via your browser’s preference settings.GeneralType: Elizabeth II Canadian Silver 25 Cents Years: 1965 - 1966 Cat. Num.: KM# 62 Country: Canada Period: Confederation, Elizabeth II Currency: Canadian Dollar Face value: 25 Cents System: Decimal 100 Cents = 1 Dollar Desgr/Engr: Obverse: Elizabeth II young bust right Legend: ELIZABETH II D·G·REGINA Reverse: Caribou left, denomination above, date at right Legend: CANADA 25 Cents Edge: Reeded Purpose: Circulation 1965 Quarter Value SilverIssueYear: Mint: Mintage: 44,708,869 Scarcity: Valuation: Notes: Grades & Prices Available (VF)Sold(EF)Sold(UNC)SoldSpecificationsComposition: Silver Fineness: 0.8000 Weight (g): 5.8319g Weight (Oz): 0.19 Oz Net Content: 0.15 Oz (4.67g) Bullion Value: $4.10 Alignment: Diameter: 23.50mm Thickness: Description
The quarter is a Canadian coin, valued at 25 cents or one-fourth of a Canadian dollar. It is a small, circular coin of silver colour. According to the Royal Canadian Mint, the official name for the coin is the 25-cent piece, but in practice it is simply called a quarter.Value RangeYearMintageGFVFEFAUUNCBUPROOF196544,708,869$4.00$4.70$5.20$7.20Similar Items in Stock$14.99$12.99$20.99$30.502008-2021 © Silveragecoins.com - Conditions of Use | Privacy | Ordering Policy | Contact UsClad CompositionMinted at: Philadelphia (No Mint Mark) Metal Composition: 91.67% Copper - 8.33% Nickel Mass / Weight: 5.67 grams What This Coin Looks Like (Obverse, Reverse, Mint Mark Location, Special Features, etc.):Coin Value Chart: Typical Coin Prices, Values and Worth in USD based on Grade/ConditionGood(G-4)Very Good(VG-8)Fine(F-12)Very Fine(VF-20)Extremely Fine(EF-40)About Uncirculated(AU-50)Uncirculated(MS-60)Uncirculated(MS-65)Proof(PR-65)-------$9.54-USA Coin Book Estimated Value of 1965 Washington Quarter is Worth $9.54 or more in Uncirculated (MS+) Mint Condition. Click here to Learn How to use Coin Price Charts. Also, click here to Learn About Grading Coins. The Melt Value shown below is how Valuable the Coin’s Metal is Worth (bare minimum value of coin). Click here to see the Melt Value of every US Coin.MELT VALUE: $0.0501Even More Washington Quarter Years/Mints/Varieties
38 items found1965 Quarter Value Worth
*The coin in the picture is the coin you will receive. See PicturesSeller: kyoung0Condition: See Pictures
*The coin in the picture is the coin you will receive. See PicturesSeller: kyoung0Condition: See Pictures
*Seller: coinratCondition: Circulated
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*The coin in the picture is the coin you will receive. See PicturesSeller: kyoung0Condition: See Pictures
*The coin in the picture is the coin you will receive. See PicturesSeller: kyoung0Condition: See Pictures
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*The coin in the picture is the coin you will receive. See PicturesSeller: kyoung0Condition: See Pictures
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*Seller: graysonhmurphyCondition: BU
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*Seller: atchisonbjCondition: MS-65 (GEM)
*Seller: GrandPawsAttic
*Seller: collectorcoins2020Condition: F-15
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*Seller: GrandPawsAtticCertification Agency: PCGSCertification Number: 5997.64/12608054Condition: MS 64 SMS
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With being at home for the next thirty days or so I thought it would be fun to create a 30-day quarantine bingo challenge. I shared it this week on my Instagram stories. I figured it would be a fun way to pass time while being somewhat productive. Work from Home Bingo makes it fun to connect and socialize with friends and coworkers. Simply download, print and play. WORDS LIST: Drinking, too, much, coffee, Junk, food, for, breakfast, Worked, from, bed, Wearing, my, pajamas, all, day, Took, shower, Used, a, zoom, background, Forgot, what, day, it, is, Slept, through, alarm, Have, 4+, conference, calls, in, a, day, Bought, a, webcam, /, printer, Longing, for, office, birthday, cake, Pile, of, dishes, on, work, desk, Kitchen, table, is.
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This bingo card has a free space and 54 words: PJs All Day, Kitchen Table Desk, Not Missing Traffic, Short Commute, dual monitors, Snuggled My Pet, My Fridge is TOO Close, Full Screen Freeze, New Co-Workers at Home, Phone Forwarding, Coffee All Day, Hand Washing, Social Distancing, Toilet Paper, Disinfectant, Soap, Miss My Best Office Buddy, Stepping Out for Fresh Air, Elbow Bump, No Handshaking, Clorox Wipes, Bleach, No March Madness, Longing for office birthday treat day, Conference Calls, Video Calls, Netflix, Lunch Break, VPN, Wireless Internet, Jabber, Shoulder Taps/Drive-bys, Alone time, Calling Help Desk, No Travel, Facetime, Business Continuity, Flexibility, Thermostat Battles, Loud Neighbors, Cancelled Vacation, No Traffic, Stay HomeStay Safe, Quarantine, Hazmat is the new Onsie, Temporarily Out of Stock, Left Something Important In The Office, Enjoying WFH, Miss The Office Snacks, Virtual Happy Hour, Can’t Wait for Happy Hour, Cozy Socks, Laugh to keep from freaking out and Only hear every fifth word but get the gist.Work From Home Bingo GameWork From Home Bingo Card GeneratorWork From Home Bingo GameMore like this:
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Greek Coin Charm Pendant- Vermeil Gold 22k Gold plated over 925 Sterling Silver, Greek Coins, Spanish Coins, Ancient Roman Coin, Medallion VermeilSupplies. 5 out of 5 stars (19,586) $ 5.00. Bestseller Favorite Add to Ancient Coin Roman Greek Byzantine Bronze silver estate lot. Greek Coins Ancient Greek coinage is known for its amazing beauty and wide variety of artistic style. Most Greek coinage were made using silver, gold, electrum (a naturally occurring alloy of silver and gold), copper alloy, and bronze. GREEK AND PROVINCIAL COIN CITIES. Wildwinds now includes the Corinth Staters special section formerly online as CorinthianStaters.com, written by John Hayes, which covers Corinthian Pegasus staters from all the issuing cities. Kyme in Aeolis 350BC EAGLE & VASE on Authentic Ancient Greek Coin i48058. EBay (highratinglowprice) Add to watchlist. Sicyonia Sicyon 350-280 BC AR. To search an expression, simply put quotation marks around it. Example: A search for ’1 franc’ is more precise than 1 franc. You may use an asterisk as a wildcard. Example: type ’5 cent.’ to find coins of 5 cents and 5 centimes. Use a dash to exclude the coins matching with a word or expression.
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South Asia, and more precisely the modern state of India has experienced the incursion of several tribes throughout its history. Many of the famous military generals of the world had made their mark in the territories of South Asia. The Greek military genius Alexander also attacked north western India albeit without much success. The invasion of Alexander took place in the year of 326 BCE. However, he succeeded in establishing several Greek colonies. He left some of his military generals and soldiers to occupy and rule his Indian and Central Asian conquests. These Greek generals came to be known as Indo-Greeks in the history. They ruled roughly during the period between mid-3rd century BCE when Diodotus I established an independent kingdom to early 1st century BCE when they were overwhelmed by the Parthians and the Shakas.
Their rule extended over a vast part of central Asia and north western South Asia. It included the modern areas of Afghanistan, north western part of Pakistan, the Indian provinces of Kashmir and Punjab. There were several dynasties and over 40 rulers of the Indo-Greek lineage who ruled over this extended time period. And surprisingly the main sources of information about the rule of these numerous kings are the numismatic evidences. Greek Coins
The Indo-Greek coins inaugurated a new phase in the history of South Asian coinage. These coins carried elaborate details about their issuing authority. The name, the issuing year of the coin and a portrait of the reigning monarch was die-struck very precisely on the metal pieces.Greek Coins Ancient
We can identify elaborate measures of coin circulation in the Indo-Greek territory. The coins circulated in the north of the Hindu Kush Mountains were mainly made of gold, silver, copper and nickel. They were struck according to Attic weight standard. The obverse of the coins carried the portrait of the issuing monarch. The reverse of the coin was marked by the depiction of Greek gods and goddesses. The name of the monarch and his royal titles were also mentioned in the reverse in Greek.
The coins which were circulated in the south of the Hindu Kush bear more Indian touch. They were mostly made of silver and copper. Most of these coins are of round shape, while some of them are square. These coins were struck according to Indian weight standard. They bear the royal portrait on the obverse. But their reverse was marked by Indian religious symbols rather than Greek. These type of coins also carried bilingual and bi-script inscriptions using the Greek and Prakrit languages; and Greek and Kharosthi or Brahmi scripts. Ancient Greek Coins
The Indo-Greek coins have been found in large numbers in the modern Afghanistan. The largest number of coins was discovered from Gardez. This hoard is known as the Mir Zakah hoard. It yielded 13,083 coins. Among these large number of coins 2,757 were Indo-Greek coins. Other major finds are the hoard found at Khisht Tepe near Qunduz and the coins found during excavations at the city of Ai-Khanoum.
The Indo-Greek coins are very important source of ancient Indian history. Out of 42 Indo-Greek kings who ruled, about 34 kings are known only through their coins. Coins of such kings as Menander depicted them slowly progressing from their teenage to old age, which also indicated their long reigns. The high standard of coinage set by the Indo-Greeks worked as a model for several other Indian dynasties for a very long period of time. The representation of Indian religious figures and symbols in the Indo-Greek coins has a greater significance for the cultural history of South Asia. This illustrated the syncretism of the Indo-Greek rulers. A sort of cultural and religious fusion between India and Greece can be traced from these coins.
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*Peter Eastgate hit an ace-to-five straight on the turn and instantly called an all-in bet from Ivan Demidov on the river to win the title and $9,152,416. Demidov held two pair, twos and fours.
*Peter Eastgate (3:11 PM) Thre have been a lot of people who are berating her for the things she did on TV. I played with her for only a little and she treated me with respect so I don’t think I.
Peter Eastgate (also known by his nicknames ’Icegate’ and ’Isser’) was born on December 13, 1985 in Odense, Denmark. He became known in the poker world for his WSOP Main Event win in 2008. Besides the bracelet and money he won, he also set a new record for becoming the youngest player to win a WSOP Main Event at the age of 22, beating Phil Hellmuth’s record which dominated the category since his win in 1989, when he was 24 years of age.
Eastgate studied at Sct. Knuds Gymnasium, where he was first exposed to the game of poker. He continued his education and went to Aarhus University, where he studied finance but decided to quit in order to play poker professionally.
Eastgate is an example for a player who made his way to the WSOP Main Event through playing online. He qualified to the event after playing in a satellite in Ladbrokes poker website and when he came to Las Vegas, Nevada to play the Main Event, he was already part of the Ladbrokes team.
Eastgate advanced to the final table with 18,375,000 chips only and was a 4-1 favorite of winning the event. With him in the final table were also Scott Montgomery with 19,690,000 in chip, Ivan Demidov with 24,400,000 in chip and Dennis Phillips with 26,295,000 in chips. At the end it was Eastgate vs. Demidov in a heads-up play for the title. The game was over when Eastgate had 120 million in chips against Demidov’s 16.45 million in chips. In this stage, Demidov decided to go all in and lost his hand to Eastgate who became the youngest champion in a WSOP Main Event.
Eastgate’s win gave him his first bracelet and an astonishing amount of $9,152,416.
Eastgate also made it to the final table in the 2007 Irish Poker Open and in the money in the 2008 European Poker Tour (EPT) event which was held in Copenhagen, Denmark. After his 2008 WSOP Main Event win, he continued his great success and went to win another event to take home $343,000. So far as of 2009, he has won over $9,700,000 in live tournaments only.
WSOP Winnings
Young Danish pro Peter Eastgate could make history at this year’s WSOP – at 22 years old, he could beat Phil Hellmuth’s record as the youngest player to win the Main Event (Hellmuth was 24 at.
2008 - $10,000 No-Limit Texas Hold’em Main Event - $9,152,416Peter Eastgate Net WorthPeter EastgateEastgate playing heads-up at the 2008 World Series of Poker Main Event.Nickname(s)Icegate, IsserResidenceCopenhagenBorn13 December 1985 (age 35)World Series of PokerBracelet(s)1Final table(s)2Money finish(es)3Highest ITMMain Event finishWinner, 2008European Poker TourTitle(s)NoneFinal table(s)2Money finish(es)7Information accurate as of 28 February 2012.
Peter Eastgate (born 13 December 1985)[1] is a poker player from Denmark, best known as the winner of the Main Event at the 2008 World Series of Poker. At the time, he became the youngest player ever to win the event.[1] He was subsequently surpassed by Joe Cada in 2009.Early life[edit]
Eastgate was raised in Dalum, a suburb of Odense, Denmark and went to Sct. Canute’s Gymnasium, where he was introduced to poker by his classmates as he stated in season 5 of the show High Stakes Poker. He attended Aarhus University to study economics, but dropped out to play poker full-time.Poker career[edit]World Series of Poker[edit]
Eastgate qualified for the Main Event via the Ladbrokes Poker website, and travelled to Las Vegas as part of Team Ladbrokes.[2] Eastgate reached the final table with 18,375,000 in starting chips, the fourth largest stack behind Scott Montgomery (19,690,000), Ivan Demidov (24,400,000) and Dennis Phillips (26,295,000).[3] Eastgate and Demidov reached the 2008 World Series of Poker (WSOP) heads-up stage. At the final hand, Eastgate had 120.4 million in chips, and Demidov went all in with 16.5 million chips. Eastgate beat Demidov at 2:33 AM local time, after a two-day final table consisting of 274 hands that took 15 hours and 28 minutes to play.[3] Eastgate won the first place prize of $9,152,416.[1][4][5]Peter Eastgate Broke
By winning the Main Event at age 22, he became the then youngest WSOP Main Event champion, surpassing Phil Hellmuth, who was 24 when he won in 1989. He was surpassed as the youngest winner of the WSOP by Joseph Cada the following year at age 21[1]
In 2009, Eastgate defended his Main Event title until day 6. He was eliminated in 78th place as a result of a rivered flush. He was the last remaining Main Event Champion at the time of his elimination.World Series of Poker braceletsYearTournamentPrize (US$)2008$10,000 World Championship No Limit Hold’em Main Event$9,119,517Other events[edit]
Prior to the WSOP, Eastgate’s live tournament results included a money finish in the 2008 European Poker Tour event in Copenhagen and a final table finish at the 2007 Irish Poker Open.[5] Eastgate’s previous biggest win was $46,714 at the 2007 Paddy Power Irish Open. After his WSOP Main Event victory, Eastgate took to the tournament trail and quickly found success. He cashed in the European Poker Tour (EPT) PokerStars Caribbean Adventure (PCA) Main Event and then won the $5,000 side event for $343,000[6] Eastgate came second in the 2009 London EPT, winning £530,000.[7]
In 2010, Eastgate finished 5th in the NBC National Heads-Up Championship in Las Vegas for $75,000.[8]
As of 2014, his total live tournament winnings exceed $11,200,000.[9]Peter Eastgate All BlackHiatus[edit]
In July 2010, Eastgate announced that he was taking a break from poker in a statement through his sponsor PokerStars.com, saying, ’When I started playing poker for a living, it was never my goal to spend the rest of my life as a professional poker player. My goal was to become financially independent. I achieved that by winning the WSOP main event in 2008. The period following has taken me on a worldwide tour, where I have seen some amazing places and met many new people; it has been a great experience.’[10]
“In the 20 months following my WSOP win, I feel that I have lost my motivation for playing high-level poker along the way, and I have decided that now is the time to find out what I want to do with the rest of my life. What this will be, I do not yet know. I have decided to take a break from live tournament poker, and try to focus on Peter Eastgate, the person. I want to thank PokerStars, my friends and family for their support over the last 20 months, and for their support in my decision to take a break from poker.’[10]
PokerStars said of Eastgate’s decision: “We know that poker will miss this world-class player and world-class role model, but PokerStars has only the deepest respect for Peter’s decision. Poker is about determination and excitement, and if Peter lacks this in his game, the right decision is to take a break. Peter has played amongst world champions and he has carried his title in the most admirable fashion.”[10]In November 2010, Eastgate put his main event bracelet up for auction on eBay with a starting bid of $16,000. It eventually sold for $147,500 [11][12] and this money was donated to UNICEF.[13]
In February 2011, Eastgate announced that he would be ending his eight-month break to return to poker.[14]Notes[edit]
*^ abcdDalla, Nolan (11 November 2008). ’Peter Eastgate Wins the 2008 World Series of Poker’. pokerplayernewspaper.com. Archived from the original on 22 April 2009. Retrieved 11 November 2008.
*^Ladbrokes congratulate Eastgate[permanent dead link], a November 2008 article from the website of Dennis Publishing’s Inside Poker magazine
*^ abGary Milhoces (11 November 2008). ’Eastgate calm as he wins World Series of Poker’. USA Today, [USA Today flash graphic]. Retrieved 12 November 2008.
*^Final nine hold ’em for now, wait until November to play, a July 2008 article from USA Today
*^ abButt, Robert. ’Peter Eastgate - Results’. TheHendonMob.com. Retrieved 11 November 2008.
*^ThePlayr Poker News - Peter Eastgate Wins EPT PCA Event
*^https://www.mirror.co.uk/poker/news/2009/10/07/ept-london-gustavson-defeats-eastgate-for-850-000-and-ept-title-115875-21730857/#
*^Hendon Mob Database: Peter Eastgate
*^Hendon Mob Database: Peter Eastgate
*^ abcMurray, Brendan (6 July 2010). ’Former World Champion Peter Eastgate Quits Poker’. CardPlayer.com. Retrieved 8 July 2010.
*^Peter Eastgate 2008 WSOP Bracelet Sold for $147,500
*^Peter Eastgate Auctions WSOP Bracelets
*^The Nightly Turbo: Peter Eastgate Auctions off WSOP Bracelet, APT Macau Crowns Winner, and More | PokerNews
*^Peter Eastgate’s return, in his own wordsExternal links[edit]Retrieved from ’https://en.wikipedia.org/w/index.php?title=Peter_Eastgate&oldid=934836228’
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