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I Miss The Mid-Missouri Outlaws [OPINION]
It has been two years since the Mid-Missouri Outlaws football team took the field at the Mathewson Exhibition Center. There were very few times that I missed a game of theirs from the hey day of when they won the outdoor leagues in the CPFL to when they joined the American Professional Football League in 2010 and made it to the championships to their final year when the APFL disbanded and the Outlaws helped start the CPIFL in 2013.
Unfortunately the last few years the team could never regain the championship form that they had in the CPFL or the fantastic start that they had in the APFL, but they all brought heart to the field. I know that the last season the team finished 0-12 and couldn't get a break against the bigger city teams and in the CPFL, teams had bigger budgets, bigger players to recruit and a lot of the owners that was their job all year around.
I loved the competition and seeing the players that would show up from other teams that I enjoyed watching in college. Some of the players around here, played for the love of the game and played like champions hardly ever hanging their head low.
I loved watching Marcus 'Superman' James run with the ball, return kickbacks and always giving high fives to kids in the end zone. He may have been the fan favorite, but who could forget when the Outlaws made ESPN with a wild and crazy play in the championship with a pass from Kyle Middleton to Jerry Young, who are Sedalia natives. Then there are players I loved to talk with, such as Tony Vaka, Mark Tiernan, Ken Davis, Tenny Barber, Derek Doerfler, Ben Lyles, Jeff Clancy, James 'Flash' Cody, Micah and Seth Brimer just to name a few that I remember of the years.
Some of them have continued on in indoor football, with a handful of them on the Kansas Koyotes now.
I think what I most miss is the entertainment on the field. The Outlaw cheerleaders had some of the craziest games from Junk in the Trunk to kicking field goals at half time for prizes to throwing footballs out after every score to of course their terrific dance routines.
I mostly loved the fierce hitting on the walls to the constant running clock. I know that the Mid-Missouri Outlaws ran into being the smallest city in a big league and that did them in, but the competition alone was worth it. Getting to see SEC, Big 12, Big 10 players all around come together.
I do wish and hope that sometime down the line, maybe we can get another indoor team in to fill our arena. Some of you may have forgot that the Missouri Minutemen played at the Mathewson from 2004-2006, before Travis Jobe and Chad Jackson put the Outlaws into the APFL a few years later. It was that taste of the Minutemen that brought the Outlaws and maybe down the line another venture could take place of some sort.
I do thank Travis Jobe, Chad Jackson and Ethan Henson for taking the chance years ago. As we know arena football is a hard one to keep up, not even the APFL is around anymore or the CPIFL, it moved to the CIF and have spread all across the Midwest. Kansas City still doesn't have an indoor team, but sometime the chance will come up, but it's at this time of the year when the leagues start playing that I really miss indoor football in Sedalia.I hope if that chance comes we get out and support it.
For fun, here is the play that brought the Outlaws national attention.


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Huttopia Divonne Huttopia
Europe France Rhône-Alpes Ain
Camping Pitch With Electricity
Mobil Home Vancouver
The Classic IV Wood & Canvas Tent
Chalet Montana
There's a bit of an irony to all the facilities at Camping Divonne Les Bains on the Swiss-French border. Set within the edges of a thick forest that tumbles down one of the highest mountains in the Jura range – La Dôle – it has a magnetic location for all outdoorsy types. Yet, geared to accommodate family campers, as well as hardy backpacking folk, there's a complete spectrum of onsite options too. A tennis court, heated outdoor swimming pool, volley ball court and pétanque pitch all leave you wondering, why the need to create such an elaborate onsite playground when the continent's natural playground is right on your doorstep?
The resulting campsite is expansive, with almost 200 different pitches. Massed among the trees, however, each pitch still retains its own sense of privacy, while multiple wash-blocks – immaculately clean and well looked after – mean, wherever you choose to pop the tent, you'll never have too far to walk. With so much on offer, there really is something for everyone and the campsite is a diverse mix of backpackers, rock climbers and mountain bikers, combined with friendly family groups, lounging by the poolside or warming up on the tennis court.
It's away from your pinecone scattered pitch that the real benefits of this campsite are found, though. Right from the entrance of the campsite there are trails leading into the trees and up the angular sides of La Dôle – a summit of 1677 meters. If you're feeling lazy, you can drive a few minutes into Switzerland and up the paved road in Gingins to Chalet de la Dôle just below the peak. If you're walking it's a great place to stop for fuel. From the top, the Alps spread away to the East, while Lake Geneva spreads below. It's here, on the waterside, that you can spend the rest of your days, boating, bathing and generally having a boon.
Video Tour
The Owner SaysChoose a nature stay at the foot of the Jura mountains, just a few kilometres from Lake Geneva and at the gateway to Switzerland!
On a 9 hectare site, tents, caravans and motorhomes can set up camp on terraced pitches, some of which offer unspoilt views of the lake and the mountains.
Under the trees or in the sunshine, there is something for everyone.
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subset: commoncrawl


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Yes, the US casinos online are legal. According to the UIGEA American law, the online gambling institutions are allowed to accept and pay out money to US players. Therefore making online gambling in the US legal. However, it is advisable that you check with the laws of your state before registering on any of the casinos online. This is to remove any risks of getting into issues with the local authority as you play online.
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There are a few common things that you should be checking out before registering yourself in any online gaming websites. They are checking if they have completely disclosed their payment procedures, policies and the rules. This will help you in knowing that there is no hidden clause that you are not aware of, that you would be threatened with later. Ensuring that the casino you are willing to register in is licensed or not is also another important step in getting the into the Top US Online Casinos. Almost all licensed casinos are generally reliable and trustworthy.
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If you search the internet, you will come up with sites that are offering reviews and ratings about some new US online casinos. The sites include what people say about it and what to consider before signing up. A lot of people seem to enjoy and are giving positive ratings to online casino site. There are just reminders to make sure the site you are signing up for is licensed and that they give out and offer fair payout percentage and RTP (Return to player) to avoid scams.
So, the only thing you need to do is to find a online casino, and try your luck to make some extra money.
Some Profitable Gambling at New Online Casinos USA Clubs
What are some of the profitable games available in New Online Casinos USA?
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If you need to pay for real money then there are some simple steps that you need to follow strictly. First you need to download the free casino software that will allow you to play online with other players elsewhere. You also need to register for a real account so that you can become a member. You also need to deposit some cash in your account and your bonus will be automatically credit to your account so that you can be in a position to play effectively with other gamblers elsewhere in different parts of the world.
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I have written a small program in assembler which works almost perfect... Almost.....
I want the program to monitor the switch at input RB0.
As soon as the button has pressed AND released, a LED at RE2 must lit for about 800mSec.
After that, the LED turns off and the program repeats.
So far, this is exactly what the program does!
- Code: Select all
;wait for button press @ RB0
banksel PORTB
wait_dn btfsc PORTB,RB0 ; skip next instruction if RB0 => LOW
goto wait_dn
;wait for button release
banksel PORTB
wait_up btfss PORTB,RB0 ; skip next instruction if RB0 => HIGH
goto wait_up
;turn on LED @ RE2
banksel LATE
bsf LATE,RE2
;delay 800mSec
movlw .80
pagesel delay10
call delay10
pagesel $
;turn off LED @ RE2
banksel LATE
bcf LATE,RE2
;repeat forever
goto MAIN_LOOP
Now, the main 'failure' of this program is that, when the LED is on, if the button is pressed within the 800mSec delay, nothing happends because the program is in the delay routine.
What I want is that if the LED is on, and if the button is pressed before the LED is turned off, the LED is switched off and the program waits unti the button is released again.
I think I know the answer to this and that is to use interrupts, right?
So, I have my switch monitoring in the MAIN_LOOP and the delay function as ISR, right?
Any tips or tricks are welcome!


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Our items for this October 15th - November 10th round of "The Garden" at TLC are out now!
This is our very first bed, and second (third if you count the treehouse) building release and I am sooooo excited!
This event is filled with some amazingly talented creators that I am super honored to participate alongside of!
For the details~
Soft Rustic Double Bed
15 Adult anims
& PG only version included
4 land impact
Tintable white base textures (5 tintable bases for the blanket)
& white base textures included for you to customise externally
It is so pretty and I am totally in love with this bed! I hope everyone will enjoy it.
Lovely Lil' Log Cabin
Kool Doors Included
Fireplace included
14 land impact for the building
2 land impact for the fireplace
16x18m required for placement, scales up wonderfully
Again! I hope you'll take the time to stop by this fabulous event and check out not only our wares, but others too!


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The Guelph Public Library offers many different devices and hardware for public use. Each item has different borrowing guidelines and can be viewed below.
In-library use
Internet computers
All locations have internet computers that can be accessed with a valid Guelph Public Library card. Visitors can use internet terminals with a visitor's pass. These computers have Microsoft office 2016, USB slots, and the ability to print. There are select computers at the Main Library that have CD drives. There are no time limits on the internet computers at the Main Library.
Catalogue computers
All locations have catalogue computers that can be used to search for library materials and to access our eResources.
The Main Library has Windows 8 laptops and Chromebooks. These laptops can be checked out at the Information desk with a valid Guelph Public Library card. You may borrow the laptops for 2 hours per day.
All branches also have Chromebooks available for loan to Upper Grand District School Board students. The loan period for these Chromebooks is 7 days. Students who wish to borrow one of these laptops require a valid Guelph Public Library card.
All branches have iPads for in-library use. The Main Library also loans Windows Surface tablets. Tablets may be loaned via the Service Desks with a valid Guelph Public Library card. Tablets may be used to access the Guelph Public Libraries WiFi and are loaned for 2 hours per day.
Printers and scanners
Printing is available at all Guelph Public Library locations. The Main Library has a scanning station set up for public use. For more information on these services check out the Printing and Copying page.
The Main Library has microfilm machines available to the public. For more information on these machines go to our Archives page.
Loanable devices
WiFi Hotspots
You may borrow a Netgear WiFi hotspot from any location to connect multiple devices to the Internet wherever you go. Netgears may be loaned for 1 week.


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Like Us on Facebook
Online Adventure Games
The online adventure games are very interesting and attractive games on the internet. Boys and girls like to play these free adventure games with heart and very excited to play new adventure games. The online adventure games are very helpful for kids in real life and motivate them to do some outdoor activities. The kids learn from these games to take risks in life and how to solve it. Free adventure games improve the skills of kids and give them different ideas. The new adventure games are developed on basis the kid's mentality. All adventure games on this page have best quality of graphics and audio. Instructions of games are given and very easy to handle and play.


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- Research article
- Open Access
The OsSec18 complex interacts with P0(P1-P2)2 to regulate vacuolar morphology in rice endosperm cell
© Sun et al.; licensee BioMed Central Ltd. 2015
- Received: 21 July 2014
- Accepted: 6 November 2014
- Published: 17 February 2015
Sec18p/N-ethylmaleimide-sensitive factor (NSF) is a conserved eukaryotic ATPase, which primarily functions in vesicle membrane fusion from yeast to human. However, the function of the OsSec18 gene, a homologue of NSF in rice, remains unknown.
In the present study, we investigated the function of OsSec18 in rice and found that OsSec18 complements the temperature-sensitive phenotype and interferes with vacuolar morphogenesis in yeast. Overexpression of OsSec18 in rice decreased the plant height and 1000-grain weight and altered the morphology of the protein bodies. Further examination revealed that OsSec18 presented as a 290-kDa complex in rice endosperm cells. Moreover, Os60sP0 was identified a component of this complex, demonstrating that the OsSec18 complex contains another complex of P0(P1-P2)2 in rice endosperm cells. Furthermore, we determined that the N-terminus of OsSec18 can interact with the N- and C-termini of Os60sP0, whereas the C-terminus of OsSec18 can only interact with the C-terminus of Os60sP0.
Our results revealed that the OsSec18 regulates vacuolar morphology in both yeast and rice endosperm cell and the OsSec18 interacts with P0(P1-P2)2 complex in rice endosperm cell.
- OsSec18
- Os60sP0(P1-P2)2 complex
- Vacuole fusion
- Rice endosperm
Sec18p/N-ethylmaleimide-sensitive factor (NSF) is a conserved ATPase required for vesicle membrane fusion in eukaryotes. In yeast and mammalian cells, the mechanism of vesicle membrane fusion, which is mediated by Sec18p/NSF and the soluble NSF attachment protein (SNAP) receptor (SNARE) complex, has been extensively investigated. NSF assembles with SNAP and SNAREs to form a 20S SNARE fusion complex that mediates membrane fusion between vesicles [1]. This 20S fusion complex is disassembled by NSF via ATP hydrolysis [2]. During this process, Sec18p/NSF, acting as a SNARE chaperone, binds to SNARE complexes, disassembling them and facilitating SNARE recycling by utilizing the energy from ATP hydrolysis. The rate of Sec18p/NSF-mediated disassembly correlates to the SNARE-activated ATPase activity of NSF [3].
NSF is also involved in protein trafficking [4-7]. Previous studies have indicated that NSF binds directly to the C-terminal tail of the GluR2 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor in a SNAP-dependent manner to regulate the function of these receptors [6,7]. McDonald et al. have found that NSF can bind to β-arrestin1 and plays a hitherto role in facilitating clathrin coat-mediated internalization of G protein-coupled receptors [8]. Cong et al. have confirmed that NSF can bind to β2 adrenergic receptors (β2-ARs) at the final three amino acids in the C-terminal tail of these receptors, thereby regulating receptor recycling [4].
To date, very limited information about Sec18p/NSF and SNARE complexes in plants is available. Sato et al. have cloned a homolog of NSF from tobacco, designated as NtNSF-1, which encodes a 739-aa protein that displays ATP binding capacity [9]. Hugueney et al. have investigated a plastid fusion and/or translocation factor (Pftf) in Capsicum annuum and demonstrated that it functions in vesicle fusion in an ATP-dependent manner. However, Pftf, which encodes a 72-kDa protein, was only expressed in leaves and young fruit in red peppers [10]. Bioinformatic analysis indicated that its cDNA sequence displays 53% and 51% homology with yeast Sec18p and mammalian NSF, respectively. However, the functions of OsSec18, a homolog of Sec18p/NSF in rice, remain unknown.
More recently, some studies have indicated that the proteins involved in protein sorting play important roles in plant development. Vacuolar protein sorting 29 (VPS29) is a component of a retromer complex that recycles the vacuolar sorting receptor VPS10 from the pre-vacuolar compartment (PVC) to the Golgi complex. In Arabidopsis, the VPS29 homolog Maigo1 (MAG1)/AtVPS29 is ubiquitously expressed in various organs, including leaves, roots, flowers and developing seeds [11]. The MAG1 mutant (mag1) exhibits a dwarf phenotype, suggesting that it may play a significant role in plant growth and development [12]. Furthermore, VPS29 is involved in endosome homeostasis, PIN protein cycling, and VSR recycling from the PVC to the trans-Golgi network (TGN) during the trafficking of soluble proteins to the lytic vacuole (LV) [13,14]. Moreover, the protein sorting protein 45 (VPS45p), a member of the Sec1p family, is involved in vesicle-mediated protein trafficking in various organelles of the endomembrane system [15,16]. Bassham et al. have found that AtVPS45p co-localized with an epidermal growth factor receptor-like protein (AtELP) in Arabidopsis in the TGN and that AtVPS45p functions in the transport of proteins to the vacuole in plants [15,16]. However, the relevance of OsSec18 and PVC remains to be determined in rice.
Ribosomal acid protein P0 as a component of P0 (P1-P2)2 complex, functioning on protein synthesis as a subunit of 60s ribosomes [17,18]. The C-terminus (199-258aa) of P0 binds to the (P1-P2) small complex [19], while the N-terminus (44-67aa) of P0 interacts to the RNA molecule after P0(P1-P2)2 complex formed [20]. Mutation of P0 gene affects the ribosome activity and viability of Saccharomyces cerevisiae [21]. Barnard et al. and Kondoh et al. have found that the human ribosomal phosphoprotein P0 may be implicated in human colorectal cancer progression [22,23]. Recently, Chang et al. have found that overexpression of P0 protein might cause oncogenesis in breast and liver tissues by partially inhibiting GCIP-mediated tumor suppression [19]. All these results suggest that P0 protein is important for the protein synthesis as well as other cellular functions, such as oncogenesis [17-19]. Rice Os60sP0 is 60% homologous to human 60sP0 in DNA sequences and 53% homologous in amino acids sequences. When compared with yeast, the homology is 54% and 46%, respectively [24]. However, the functions of P0 (P1-P2)2 complex in rice have not been previously reported.
In the present study, we investigated the function of OsSec18 in rice and found that it can complement the temperature-sensitive phenotype but cannot restore vacuolar morphology in yeast. This result suggests that the OsSec18 gene may perform other unknown functions than in yeast. Overexpression of the OsSec18 gene in rice decreased the plant height and 1000-grain weight, and changed the morphology of the protein bodies. Further studies demonstrated that OsSec18 is a component of a 290-kDa complex in rice endosperm cells. Moreover, Os60sP0 was identified as a component of this complex, revealing that the OsSec18 complex contains another complex of P0(P1-P2)2 in rice endosperm cells. Furthermore, we determined that the N-terminus of OsSec18 interacts with the N- and C-termini of Os60sP0, whereas the C-terminus of OsSec18 interacts only with the C-terminus of Os60sP0. We proposed a molecular model for the interaction between OsSec18 and Os60sP0.
The expression profile of OsSEC18 in rice
Although Sec18 has been extensively studied in yeast and mammals, its functions in plants remain unknown. To investigate the function of Sec18 in rice, we first searched the rice genome database (www.gramene.org). An OsSec18 gene (GenBank No. Os05g0519400) is homologous to SEC18 in yeast. OsSec18 shares 46%, 45%, 75% and 37% homology with tobacco NSF, yeast Sec18p, human NtNSF-1 and Capsicum annuum Pftf, respectively (Additional file 1: Figure S1 and Additional file 2: Figure S2). OsSec18 contains two AAA ATP domains at the C terminus and the middle region of the amino acids sequence, and it displays ATP-binding and nucleotide-binding nucleoside-triphosphatase activity.
Interestingly, we found three isoforms or modifications of OsSec18. OsSec18 displayed the lowest molecular mass in inflorescence and immature seed, followed by mature seed and stem, and the highest mass in leaf. These results indicated that OsSec18 is expressed as distinct isoforms or is modified in a tissue-specific manner, implying that these isoforms or modifications may play distinct roles in different organs or tissues.
OsSec18 does not completely complement the function of vesicle fusion in the yeast sec18 mutant
Yeast complementation assays
Selective medium
Growth temperature
Clearly, vesicle fusion was disturbed when OsSec18 was expressed in yeast cells. These results showed that the OsSec18 gene not only restored the ability of sey5186 to grow at 37°C but also interfered with vesicle fusion, thus altering vacuolar morphology in yeast. This result suggests that OsSec18 performs nearly the same growth-related function as Sec18/NSF in yeast, but OsSec18 also interrupts vesicle fusion and vacuolar morphology.
Overexpression of OsSec18 alters the morphology of the protein bodies
Phenotypic analyses of transgenic and wild-type rice
Plant height (cm)
+/− (%)
1000-grain weight (g)
+/− (%)
OsSec18 is a component of a 290-kDa complex in rice endosperm cells
P0(P1-P2)2 is a component of the OsSec18 complex in vivo
Although Sec18p has been extensively studied in yeast and mammalian cells, its functions in plant vacuolar compartments remain to be determined. In this study, we found that OsSec18 rescued a yeast temperature-sensitive mutant phenotype and affected vacuole morphology by interfering with vesicle fusion when overexpressed in either mutant sey52186 or wild-type sey6120 cells in yeast. Three isoforms of OsSec18 were found in different tissues of rice. Furthermore, our data further indicated that OsSec18 affects the morphology of PSV in rice endosperm cells. Moreover, we identified a 290-kDa complex of OsSec18 in rice endosperm and demonstrated that heterologous complex P0(P1-P2)2 is another component of OsSec18 complex. Our data indicate that OsSec18, along with heterologous complex P0(P1-P2)2, is involved in rice vacuolar morphogenesis.
Recently, Jaillais et al. studied vacuolar protein sorting 29 (VPS29), a single-copy gene in Arabidopsis that is an ortholog of VPS29 in yeast and mammals [13]. They found that not only is the AtVPS29 protein a member of the retromer complex but also is required for endosome homeostasis, PIN protein cycling and dynamic PIN1 repolarization during development. Although the interaction among OsSec18, Os60sP0 and PVC had not been reported previously, several studies indicated that the novel function of VPS genes rather than vacuolar fusion and protein trafficking [13,16] . In our study, we found that Os60sP0 play a novel function of vacuolar morphology than protein synthesis. In general, ribosomal acid protein P0, a component of the P0(P1-P2)2 complex, is a subunit of the 60s ribosomal complex that mediates protein synthesis [17,18]. Previous studies indicated that mutation of the P0 gene affects ribosome activity and cell viability in Saccharomyces cerevisiae [21]. Barnard et al. have found that human ribosomal protein P0 phosphorylation is involved in the progression and biological aggressiveness of human colorectal cancer [22]. Furthermore, Kondoh et al. found that P0 may exert a causal effect on hepatocellular carcinoma (HCC) progression via the translational machinery due to its interaction with eukaryotic elongation factors [23]. Recently, Chang et al. reported that the overexpression of the P0 protein may cause tumorigenesis in breast and liver tissues, which at least partially inhibited GCIP-mediated tumor suppression [19]. Based on our data, we found that OsSec18 interacted with P0(P1-P2)2 to form an OsSec18-P0(P1-P2)2 complex. Serial deletion mutation demonstrated that OsSec18 binds to the Os60sP0p in a head/tail to head manner. Our findings provide insights into the functions of OsSec18 in plant growth, vesicle fusion and vacuolar morphology.
In our study, we found three isoforms of OsSec18 in different tissues, clearly suggesting that each isoform may have a unique function in each tissue. The isoform with the highest molecular mass was expressed in leaves, whereas the vacuole morphology and function are largely different from those of other tissues. In addition, the middle size isoform was found in stems and mature seeds, where the vacuoles are transformed into storage- or transportation-related organelles. The smallest isoform was found in inflorescences and immature seeds, which have highly active sites of protein synthesis and cell division. However, the mechanisms by which these isoforms are formed remain unknown. Several mechanisms could underlie the formation of these isoforms. One possible mechanism is alternative splicing at the transcriptional level; another possible mechanism is post-translational modification such as phosphorylation. Thus, our findings introduce new avenues of investigation into the functions of vacuolar fusion in higher plants. It will be interesting to explore the functions of different isoforms of OsSec18 in rice in future.
In the present study, we found that OsSec18 is a component of a 290-kDa complex in rice endosperm cells, and moreover, Os60sP0 was identified as a component of this complex, revealing that the OsSec18 complex contains another complex of P0(P1-P2)2 in rice endosperm cells. Furthermore, we determined that the N-terminus of OsSec18 interacts with the N- and C-termini of Os60sP0, whereas the C-terminus of OsSec18 interacts only with the C-terminus of Os60sP0. We propose a molecular model for the interaction between OsSec18 and Os60sP0.
The S. cerevisiae sec18 mutant sey5186, carrying the genotype MAT sec18-1 ura3-52 leu2-3, 112 GAL + , and wild-type sey6210, carrying the genotype MAT ura3-52 leu2-3,112 his3-200 trp1-901 lys2-801 suc2-9, were kindly provided by Karl Fu. The OsSec18 cDNA clone was purchased from Japanese NIAS GenBank (Accession No. AK072976). A japonica variety, TP309, was used in all plant experiments. All biological reagents, including enzymes, kits, and biomaterials, are listed in Additional file 3: Table S1.
Genetic complementation assays in yeast
A full-length cDNA of rice Sec18 gene was digested by restriction enzymes, SacI and BamHI, and then inserted into the pYES.2 vector (Invitrogen, Carlsbad, CA, USA) and designated as pOsPMP77. The plasmid was introduced into the sec18p mutant strain Sey5186 and the wild-type strain Sey6210, following standard protocols [26]. The transformant strains were grown at 37°C for 30 hrs. The sample preparation for electron microscopy was performed as described by Yang et al. [27]. The colony phenotypes and cellular microstructures were observed via transmission electron microscopy (FEI Company, Hillsboro, OR, USA).
Plasmid construction and transgenic plant generation
An overexpression vector driven by the CaMV35S promoter was constructed. Briefly, the full-length cDNA encoding OsSec18 (GenBank accession No. J023146P19) was digested using the restriction enzymes BamHI and EcoRI and then inserted into the PKANNIBLE plasmid vector. The resulting plasmid was designated as pOsPMP124. Transgenic plants containing the overexpression plasmids were generated via biolistic bombardment-mediated transformation.
Antiserum preparations
A serum against OsSec18 was prepared by Shanghai ImmunoGen Biological Technology. The sera against Os60sP0p and Os60sP1p were prepared by Nanjing Genscript Company. Briefly, the full-length OsSec18 cDNA was inserted into the pET32a plasmid for fusion with a His tag. The engineered E. coli strain BL21 was incubated at 30°C for 6 hrs after induction using IPTG. After harvesting the cells, crude protein was extracted in phosphate buffered saline (PBS). After clarification via centrifugation at 6000 g at 4°C, the crude protein was purified using a His-tagged affinity column. The full-length OsSec18 was used as the antigen to inoculate rabbits; these antibodies were generated by Shanghai ImmunoGen Biological Technology (Shanghai, China). For the preparation of antibodies against Os60sP0p and Os60sP1p, the appropriate peptides derived from Os60sP0p and Os60sP1p were synthesized and used as antigens for inoculation of rabbits; these antibodies were prepared by Genscript Company (Nanjing, China).
SEC and MADLI-TOF mass spectrometry analyses
Rice immature endosperm was harvested after 10–14 days of pollination. Total soluble protein was extracted using PBS containing 0.1% MG132 and 1% protease inhibitor cocktail (Sigma Aldrich, St. Louis, MO, USA). The crude protein extracts were clarified via centrifugation at 10,000 g at 4°C for 10 min. The total protein was filtered through a 0.8-μm filter (Millipore, Billerica, MA, USA). The protein solution was loaded on a Sepharose 300 column (GE Healthcare, Fairfield, CT, USA) and collected in fractions of 2 mL/tube. The protein fractions were separated via 10% SDS-PAGE and then analyzed via Western blot using the appropriate antisera described above. The corresponding proteins and the appropriate molecular mass markers were separated via SDS-PAGE. The proteins of interest were carefully excised following SDS-PAGE and were washed with buffer I (50% v/v acetonitrile, 100 mM ammonium bicarbonate, pH 8.0) and incubated in buffer II (10 mM DTT in 50 mM ammonium bicarbonate, pH 8.0) at 65°C for 1 h, followed by incubation in buffer III (55 mM iodine ammonium acetate, 50 mM ammonium bicarbonate, pH 8.0) in a dark room for 30 min. Enzymatic hydrolysis was performed using trypsin (Promega, Madison, WI, USA) at 37°C overnight after washing with 10 mM ammonium bicarbonate and acetonitrile. An equal volume of buffer IV (60% v/v acetonitrile, 5% formic acid) was added, and then, the sample was ultrasonicated for 10 min. The supernatant was vacuumed dry, and 3 μL of buffer IV was added to dissolve the protein pellets. The resulting protein fractions were used for MADLI-TOF mass spectrometry analysis. The mass spectra were recorded using an Ettan MALDI-TOF/Pro mass spectrometer (ABI, Carlsbad, CA, USA), and the MS data were analyzed using Scaffold software.
Yeast two-hybrid analysis
A yeast two-hybrid kit was used. Briefly, the full-length and various deletion constructs of the OsSec18 and Os60sP0 genes were inserted into pGBKT7 and pGADT7 (Clontech, Mountain View, CA, USA) according to the manufacturer's instructions, and the constructed plasmids were transformed into the yeast strain AH109 using the LiAc method [28]. The yeast strains were grown in YPDA media (20 g/L tryptophan, 10 g/L yeast extract, 0.03 g/L adenine, 50 mL 40% glucose, 20 g/L agar, pH 5.8) or SD media (Shanghai Genomics, Shanghai, China) at 30°C or 25°C. The transformants were grown on SD medium lacking leucine and tryptophan, on SD medium lacking leucine, tryptophan and histidine or on SD medium lacking leucine, tryptophan, histidine and adenine. A yeast strain co-transformed with pGBKT7-p53 and pGADT7-T was used as a positive control, and a yeast strain co-transformed with pGBKT7-lam and pGADT7-T was used as a negative control.
Co-IP and Western blot analysis
The crude protein extracts from immature endosperm (approximately 200 mg) was obtained using 2 mL RIPA buffer (PBS containing 0.1% MG132 and 1% protease inhibitor cocktail, Sigma Aldrich, St. Louis, MO, USA) and then centrifuged at 12,000 g for 10 min at 4°C. The crude protein extracts were pre-precipitated using 80 μL of Protein A agarose beads (Beyotime, Shanghai, China) at 4°C for 1 h. The supernatant was collected via brief centrifugation, and antiserum against OsSec18 or Os60sP0p was added to the supernatant and then rotated at 4°C for 2 h. Then, 60 μL of Protein A agarose beads was added to the supernatant, and the samples were rotated again at 4°C for 1 h. The precipitated protein complexes were separated via centrifugation at 12,000 g at 4°C for 2 min and then washed three times with 1 mL of PBS containing 1 mM PMSF. Then, 20 μL of total protein extraction buffer (66 mM Tris–HCl, pH 6.8, 2% SDS, 2% β-ME) was added to dissolve the pellets. The sample was separated via 12% SDS-PAGE, followed by Western blot analysis using antiserum against OsSec18, Os60sP0p or Os60sP1p. RIPA buffer was used as a negative control for this experiment.
Electron microscopy
The sample preparation and observation for electron microscopy were performed as described by Yang et al. [27].
We thank Karl Fu for kindly providing the S. cerevisiae Sec18 mutant strain sey5186 and the wild-type strain sey6210. This study was funded by the National Foundation of Sciences (3087496).
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This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.


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Sirène Body Lotion
"The sea, once it casts its spell, holds one in its net of wonder forever."
- Jacques Yves Cousteau
This scent is light, yet sensuous. Mineral rich Sirene has notes of sweet berries with just a hint of rose. Warmed by coconut and cloves, it is rounded out with bare traces of amber and sandalwood.


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If the structure of human mind remain unchanged, we will always end up re-creating fundamentally the same world, with the same issues
BioHarmonizing '17 eCourse
BioHarmonizing 2017
Hello! Thanks for connecting! I am Frank. Dharma instructor, facilitator of the Eckhart Tolle Vancouver meetup, traveler. Being in becoming.
Here, you find inspirations about cultivating joyful living for the benefit of all beings. Please, take a look at book "BioHarmonizing: How To Flourish And Cultivate Peace" and eCourse "BioHarmonizing 2013". THANKS!


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Tag: Yandina
The Shared • Sunday 15th November
We're stoked to be playing one of The Shared's ROOTS ROCK REGGAE Sunday Soirees.
2-7 pm Sunday 15th November $12 / $10 (Friends of The Shared) @ the Door
The Flumes are on at 4pm sharp so get there early for a big arvo of cool tunes and of course the warm Caribbean hospitality and funky space The Shared crew always lay on.
YANDINA STREET FAIR • Sunday 23rd August
The 38th Yandina Street Fair! taking over the village from 11am Sunday 23rd August. Find all the info here
YANDINA STREET FAIR – Sunday 17th August
We're playing the 37th Yandina Street Fair this Sunday 17th August. It's our 1st show after a month touring the waterfalls of the deep north, Mr. Brett Lanes will be joining us on skins, and we can't wait!
The Shared
We were extremely pleased to have been invited to play a sneaky duo set at the quite frankly incredible Sunshine Coast venue The Shared. Go and see Colin Beard's current show "Icons of Rock'n'Roll" (Yes, that is the Colin Beard).
The Shared is a sanctuary, a home-away-from-home for Sunshine Coast creatives. Continue reading "The Shared"


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With banks and other lenders tightening their belts on lending criteria, Global Lender Equities First Holdings is seeing a growing trend in margin a stock-based loans. Getting a large loan for capitol or for personal reasons is not as easy as it once was due to uneasy global and national economic climates. For those who need to raise capitol quickly or do not qualify for a conventional loan based on credit, equities lending is becoming a popular alternative.
The appeal of stock-based loans
While traditional options for borrowing money still remain for many individuals seeking capitol and large loans, many banks have made it more difficult to qualify for a loan by tightening qualifications and increasing interest rates. Loans that are collateralized by stocks is a unique and attractive alternative for borrowers to receive the capitol they need. With a higher loan-to-value ratio than margin loans, stock-based loans are attractive to borrowers because they provide certainty throughout the life of the transaction and offer a fixed interest rate. Also, a stock-based loan offers a hedge that can lower the borrower's investment risk in downside market.
The innovative thinking of Equities First Holdings
Offering fresh, innovative, and alternative lending options is what Equities First Holdings is all about.
About Equities First Holdings
Equities First Holdings has 14 years of operation under its belt with offices in London, Indianapolis, Hong Kong, Singapore, Bangkok, Sydney and Perth and is a global leader in shareholder finance. Since its establishment in 2002, EFH has experienced some amazing growth. Its closed loans transactions increased 30% yearly in the past 3 years and the firm's total workforce grew by more than 50% since 2012. To support the firms continued growth and success, Equities First Holdings announced its acquisition of Meridian Equity Partners Limited in the United Kingdom. President and founder, Al Christy, Jr., believes the acquisition will bring EFH to a new and higher level of global of success.


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Meetings meetings meetings
The SkyLab team just wrapped up 51/2 hours of of meetings…*whew*. After briefly going over this week's progress, we discussed a myriad of decisions ranging from game design, to art direction, to tools implementation.
We solidified several design decisions that were sort of up in the air, we concretely determined what pieces of beta feedback we would take into account, and we talked about how all these choices would inform the art direction (and how the art direction should inform these choices). The new art style is still in it's infancy and undergoing rapid iteration, so we can't announce or show anything just yet (sorry!) but we're making progress.
New Programmer!
Although he's already been with us for a week, we're excited to officially announce the signing of Tyler Hiemke to the SkyLab development team! He joins us with extensive indie development experience creating the world-building tool Caeludum.
With Tyler, we discussed how to go about implementing a better automated maze generation tool. Our previous tool was text based – take a text file representation of a maze (autogenerated by script, modified by hand) and build the maze in Unity. With Tyler's expertise, we're going to build an interactive in-editor tool that allows easier and faster level design changes, while still giving us the time-saving benefit of automated generation.
Tyler will also be helping us support a new tile-based grid layout, which I'll be implementing to create a 3-lane movement system (one of the more common pieces of feedback we got, and just one of many design decisions we made today).
Why do I care?
What this means for those reading, is that the next beta release (0.4) will not be out anytime soon. Instead of pushing for weekly release cycles like the past 3 versions, we plan on taking the next month to implement all of these design choices and new art direction before returning to the more rapid weekly cycle. So, see ya in a month!


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DC-Unlocker Software v.1.0032 is out!
Visit DC-Unlocker Product Page
DC-Unlocker Software v.1.0032 is out! Huawei CDMA Modems Unlocker client now features Unlock for Huawei EC1561/EC152 and more!
New in DC-Unlocker Software v.1.0032:
- Added support for the following Huawei models:
- EC1561 - added Unlock
- EC152 - added Unlock
- Repair MEID or (and) ESN
- Automatically generated (and displayed before writing) pseudo ESN
- Write MEID if not present


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An album of team pictures for Rugby. To see the pictures, click on this link. Flickr album: Falcon Rugby Teams. This album will open in a new window.
There is also an album of pictures taken 'in play' at matches. Here is the link for that Flickr album: Falcon Rugby Matches. Again, this will open in a new window.
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One of the mysteries of the English language finally explained.
What is the origin of the word 'hazard'?
The word hazard, meaning 'a danger or risk' is taken from Old French, although the origin of the French word is uncertain. It is potentially of Arabic origin as, according to William of Tyre (the medieval chronicler), the same name was given to a dice game allegedly invented during the siege of a castle called Hasart or Asart in Palestine, and thus named after it, although early evidence for this sense is not substantial.
Back to word origins.
You may also be interested in
What is the origin of the word 'berserk'?
What is the origin of the word 'ye'?
What's the origin of the phrase 'to have an albatross around one's neck'?
See more from Word origins
Further reading
Top tips for CV writingRead more
In this article we explore how to impress employers with a spot-on CV.


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WELCOME to the future of transport - the UK's first road-legal solar car.
And one young man from York is part of the team that designed it.
Bootham School old boy Anthony Law worked on the project while studying engineering at the Cambridge University.
As part of the university's Eco Racing team, the 22-year-old joined other students in building the solar-powered racing car to enter in the World Solar Challenge in 2008. And he brought it to Bootham School earlier this week.
"We are driving our prototype vehicle from Land's End to John O'Groats as part of our outreach activities," he said.
"We have a dedicated and fully-trained team of volunteers - myself included - and we hope that by showing young people our solar car, with its exciting and futuristic design, we can educate and inspire them about environmental issues and sustainable technologies."
He said it was vital to look at alternative, sustainable technologies because road travel contributed more than 20 per cent of the UK's CO2 emissions.
He said: "Our solar-powered car, Affinity, shows that sustainable transport can be glamorous - not only does Affinity have infinite range and zero emissions, it also looks amazing."


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In the latest edition of Frontiers in Finance, we explore some current themes and address the opportunities digitalization presents to financial firms such as yours.
How is digital labor influencing the role of your human workforce?
There have long been warnings that robots will take over the workplace. This is no science fiction. There is a real concern that automation will make the human workforce redundant – literally. Yet, digital labor also means humans can avoid the mundane, repetitive tasks. Instead, they can be free for more value-adding roles such as strategic thinking or enhancing customer relationships.
(click here to enlarge the graphic)
If you want to see your organizations thrive in the newly automated future, then craft a detailed plan that can help assess and prepare for the impact digital labor may have on your workforces:
- Translating business strategies into people implications: Think about where you are headed as an organization and how cognitive technologies can help execute that strategy.
- Shaping and designing the future workforce: Explore different scenarios that might impact the organization and develop apprioriate responses to the most likely ones; create detailed blueprints of how human and digital labor can be optimally integrated across the organization.
- Facilitate change: Create and follow a strategic plan to help identify new job roles, and to begin training existing staff to fill newly created positions.
- Monitor progress: Adpot an agile response to ensure all risks are managed, including the supply of talented and capable people
How should you bridge the gap between financial regulation and technology innovation?
If you are concerned about how to integrate automated business processes, you are not alone. The vast majority of CEOs across industries feel the same, particularly when it comes to artificial intelligence and cognitive processes.
Regtech (regulatory technology) is uniquely positioned to assist companies to not only control costs and manage regulatory requirements, but also to address other critical areas that can help impove customer service, develop new offerings and achieve greater competitive differentiation. While many financial services firms are already investing in substantial legacy system transformations, regtech solution implemented in key areas can have a further transformative impact on an organization's technological readiness.
The rapid rise of insurtech investment
Digitalization is continuously evolving and changing shape, and is attracting huge development investment. A reflection of the growing significance being placed on industry transformation has been the rapid rise in venture capital investment in insurtech globally. As insurtech companies mature and show success, investments in insurtech will likely continue to grow. Keeping abreast of evolving trends in insurtech is critical to the long-term ability of insurance companies to compete.
Is digitalization an opportunity or a threat?
Put simply, it is both. No financial institution is immune from digitalization's charms or its threats. There has never been a better opportunity for companies to innovate and reinvent themselves at previously unimaginable speed. And yet it could also be seen as the worst of times – as we look at our roadmap to remaining relevant and sustainable in the digital age.
Full publication: Frontiers in Finance
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Girl yes! I'm obsessed with this. I've been trying to wear more minimalist outfits lately because they always look so clean and chic. Love this.
I love this chic and simple look! I love your slide shoes too! xo Jana | http://www.janastyleblog.com
Ok those shoes are way cute!!!
That's such a cute tee! It looks so comfortable and cute!
Casual chic for the win! Love it! https://fash-n-curious.com/
Need this knot tee! You can never have too many. 😉
I love how this tee just has the right detail to make it super stylish, xo
This is such a good look!! I feel like you can dress it up or down! Also, obsessed with your shoes! I've always wanted to get those but just haven't had the time!!
xoxo, sindy spencer –https://sindyspencer.com
Nice Outfit..


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Feb. 2011
As 2011
is in full swing, we wanted to tell you where we are and our plans for the
Firstly, a note from Jen:
Hi Folks,
I wanted to tell you that I have retired from GEG. This was a difficult decision, and one certainly not made lightly, but it is the right course.
I have been so fortunate and privileged to have worked with Dwayne, Eric, Brett & Paul, and they are like my brothers. It was amazing to be able to go to work with some of my favorite people - laughing, sharing, and feeling so much together. Knowing each of them has been such a blessing.
I want to thank all of you folks for sharing
your time, as you've been an integral part of what has made this so special. I wish you all abundant joy, peace, and good
fortune, and hope to see you at a GEG show in the very near future!
Thank You!
While we are sad to see our 'sister' leave the realm of GEG, we are truly happy for her new path. Her words are mutually shared by all of us.
We are
also excited, though for what the future brings! We are recording and writing again and are
working on that now. Also, we will be
playing live April 2 at Tin Angel in
Dwayne, Eric, Brett, and Paul.


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Our club was formed to meet the needs of West Maui by providing services that directly benefit our local community. We are committed to raising awareness of blindness, hearing and diabetes and their prevention
Recent Events 'TEEN DANCE' -September 2, 2012
West Maui Lions hosted a Teen Dance Scholarship Fundraiser, at Lulu's Surf and Grill on September 2nd, 2012, from 8:00pm-11:00pm. The teens had a great time enjoying Lion DJ Ron, who wow'd them with music and mixes. The B-Boys dances were also awesome and kept the teens interest as they took over the dance floor. There were about 13 Lions that participated in making sure the teens had a fun, enjoyable and safe event. We look forward to our next Teen dance on October 7th at Lulu's. Spread the word and come out and enjoy good dancing and good food from Lulu's Surf and Grill. Lulu's is also the location of our monthly General Meeting, so if anyone is interested in becoming a member, please contact Filipina Flores at firstname.lastname@example.com or email our secretary Linda Elms at email@example.com, or any member, including our President Don Young at firstname.lastname@example.com. We will also be supporting the Boy's and Girls Club of Lahaina and their Director and our Lions Danae Marin. Other organizations such as Teen Straight Talk, Director/Founder Mary Duke, is also one of our Lions who minister minister to Youth all over the world.
West Maui Lions photo's taken at the beginning our our Teen Dance, with our President Don Young
West Maui Lions President Don Young, Zone Chair Dennis Elms, and Lions Bobi, Shannin and Ethel at the Teen Dance.
West Maui's newest Lion Danae Marin, Director of Boys and Girls Club, and we are so happy to have her as one of our members. Please support the Boy's and Girls Club in any way possible and listen up for forthcoming events.
Teenagers at the Teen Dance, two awesome young ladies
Three Teenager at the Teen Dance enjoying the food and event.
West Maui Lions DJ Ron Tolbert, a talented man with lots of enthusiasm, great to have him as part of our team.
Teenager at the Teen Dance enjoying the event.
West Maui Lions Mel and Bobi, two dedicated individuals who we are proud to have as our club members.
Lulu's very own DJ Mike and Lion Barbara (firstname.lastname@example.com. Mike supported the West Maui Lions and we certainly appreciate him staying with us throughout the event.
West Maui Lions Treasurer 'Lateah Hawkins' email@example.com
Zone Chair Dennis Elms and Lion Amina Oleiwan, who is responsible for the upkeep of our Facebook page, and also fitness for lions.
CAR WASH FUNDRAISER- August 25, 2012
West Maui Lions president Don Young at the car wash standing next to our sign that was designed by Lion Arnetta Stokes. If anyone is interested in having a sign made, please contact Linda Elms at firstname.lastname@example.com
Desiree and Rachel at the Car Wash, support of West Maui Lions. Thanks for all you do and your support!! Desiree is the daughter of our President Don Young.
Lions hard at work during the car wash.
Our future Leo's at the car wash in charge of the water and soda. Grandchildren of Lion Don and Lion Dennis and Linda. Children of Lions Barbara and Desiree. We encourage our children to help and allow them to get involved with our service projects to prepare them for the future of lionism.
Our lions at work!!
The West Maui Lions Club will have it's 4th Annual Slipper Drive on July 21st, from 10:00am-2:00pm in front of Longs at the Cannery Mall in Lahaina. All slippers will be distributed to the West Maui Elementary schools to distribute to students who are in need of slippers. This year we are expecting over 400 pairs of slippers and have already started receiving donations from our mainland supporter Walter Paladino, M.D. , from Warren, Ohio who contributed $400.00 towards this years slipper drive.
Mary Duke of Teen Straight Talk who is also a member of our West Maui Lions club and surrounding communities in Ohio, are also collecting slippers to ship to Maui in support of our drive. All Lions clubs of Maui will also drop off slippers and donations. We want this to be a huge success and one of our largest slipper drives, so we need support from our community. Please stop by and drop off slippers or make a contribution so we can make sure our children are taken care of should they break their slippers during school or need slippers to wear. For further questions, please feel free to contact Linda Elms, secretary West Maui Lions Club at 276-1191.
Support Teen Straight Talk who are supporters and members of West Maui Lions. see below logo at http://teenstraighttalk.com/48
Recent Events
Joint Installation/New Member Induction
West Maui Lions clubs attended the joint installation on June 26th at the ILWU in Wailuku. There were 16 of our Lions present for the installation of which 8 members were new and received their new member certificates and emblems. We were touched as they smiled so hard with excitement of being members of our club. New members who were able to attend were; Bobby, Jennifer, Ron, Mel, Jabari, Wynne, Rossella, Mariam. We were also proud to have our newly elected officers installed; Donald Young-President, Linda Elms -Secretary and Lateah Hawkins -Treasurer, other elected officers had to work, so were not able to attend. We were also graced by our Past President-Charles Cook as he sang the National Anthem.
New members not present for the joint installation are: Edna Bush, Lawrence Bush, Mary Duke, Crystal Duke, Christopher Cook, Miranda Cook, Loreatha Hawkins, Leslie Hawkins, Lee Tillis, Audrey Tillis, Arnetta Stokes and Ashley Stokes, of which are on the mainland, but glad to be our club members. regular members, Barbara Smith, Charles Cook, Lateah Hawkins, Cecilia Pantanilla, Carmen Oleiwan, Amina Oleiwan, Filipina Flores, Wynne Simplot, Linda Elms, Dennis Elms. Also one of our soon to be lions Shirley Holmstrom who is waiting to attend her first meeting.
Congratulations to Mary Duke, who has attended some of the Lions clubs in Ohio, and is also a future guest speaker at their club. Mary and her daughter Crystal are both enjoying them roles as lions and are an asset to our club. Mary and Crystal 'You ROAR'.
Our newly elected President Donald Young, who we are all proud of will lead us into another exciting and memorable year of membership excellence and service throughout our community. We look forward to his leadership and know he will be an outstanding president for our club. Don 'You ROAR"
West Maui Lions club adopted mile marker 18-20 on Honoapiilani Highway, and starting cleaning and bagging trash at 7:30am. With the support of 9 lions members of our club we were able to bag approximately 27 large bags of trash. After completion of our activity, we were so proud to see how nice our highway looked and even members from the community commended us as we walked picking up trash by blowing their horns and thanking us. Follow we had breakfast at our new McDonald's in Lahaina, and met one of our disabled death members, who was in need of food, and didn't have money. Lion Barbara who know signing, bought him breakfast and he joined us as we ate and talked story with him and each other. 14 of our members had to work, but those who were able to come really had a great time working towards a great cause.
Scholarship Fundraiser- Held Annually At The Ritz-Carlton, Kapalua
Start planning and raising funds now to we wont' have any problem reaching our goal of $5,000.00. All proceeds raised towards this cause are allocated to Lahainaluna School ad Maui Preparatory Academy, both in Lahaina, Maui. Thru the help of the community were were able to raise $3,400.00 last year, so we would like to double that amount this year or even higher. See photos from 2012 scholarship fundraser.
September 25th - Adopt A Highway Cleanup
September 17th - Slipper Drive Drop-Off at West Maui Elementary Schools
August 28th - Car Wash - Mahalo to all who came out in support!
Lions Clubs International is the world's largest service club organization with more than 1.4 million members in approximately 46,000 clubs in more than 200 countries and geographical areas around the world.


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Will of Thomas Gushue
In re
In the name of God Amen. The thirteenth day of April one thousand eight hundred and forty. I Thomas gushue of Brigus Conception Bay, Planter being very sick and weak of body (and being about to undergo a severe surgical operation) but of perfect mind and memory thanks be given unto God, therefore calling unto mind the mortality of my body, knowing that it is appointed once unto all men to die do make and ordain this my last will and testament, that is to say, principally and first of all, I give and recommend my soul into the hands of Almighty God that gave it and my body I recommend to the earth to be buried in decent Christian burial at the discretion of my executors nothing doubting but at the general resurrection I shall receive the same again by the almighty power of God. And as touching such worldly estate wherewith it has pleased God to bless to me in this life I give demise and dispose of the same in the following manner and form. First I give and bequeath unto Amy my dearly beloved wife the sum of forty pounds current money of this Island p annum to be raised and levied out of my estate until my son Charles Gushue attains the full age of twenty one years and after that it is to be reduced to thirty pounds currency pr annum until my youngest son George Gushue attains the age of twenty one at which time the monies then remaining to be equally divided between my three sons or so many of them as may be then living. I likewise do give unto my beloved wife all my property household furniture and everything that belongs to me to have to her own use for the support of herself & family as long as she lives and at her decease to be equally divided between my three sons or so many of them as may be then living but should the said Amy Gushue marry again then in that case the sums of money before named are to be stopped from her and given for the sole support of my own children at the discretion of my executor and the property let out for their use untill the youngest son attains the age of twenty one years at which time it is to be divided as before named. Also I give to my son Charles Gushue my Gun and to my son George Gushue my watch. The necessary expenses attending my interment to be paid by my executors out of my estate and my wish is to have a neat stone to record the date of my death &c. to be chosen by my executors. Also I do appoint Mr. John Leamon and Mr. John Percey, Edward's son, to be my sole executor of this my last will and testament, and I do hereby utterly disallow, revoke and disannul all and every other former testaments wills legacies bequests and executors by me in any way before named willed and bequeathed ratifying and confirming this and no other to be my last will and testament.
Certified correct,
|Note: The wills in those will books are NOT actual wills. They are either hand-written copies or in later years typed copies of a, "last will and testament," written by the court clerk, after the death of the testator, when the executor presented them to the court for probate. The court clerk didn't list the signatures at the bottom, he (or she) just put them in the book in whatever order they were in, on the original document, no spacing most of the time, no punctuation. The originals were kept by the executor.
We who have typed these wills, have made every effort to include all the errors that were on the microfilm, in order to avoid destroying the integrity of the originals, where ever they may be.
Page Contributed by Judy Benson & Ivy F. Benoit
Page Revised by Ivy F. Benoit (Wednesday February 20, 2013 AST)
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Silver Tree Residential is Investing Heavily in Gambrill Gardens to set the Standard of Senior Living in Ellisville.
*Estimate Completion Date of 1/15/18*
Today's senior living marketplace is a dynamic and fast changing arena. The U.S. population over age 65 is growing steadily and senior communities that cater to this population must offer modern amenities in a stylish environment.
Gambrill Gardens will boast upscale aesthetics coupled with a hospitality-style environment. A place where seniors can age in place, with needed care at their fingertips in an engaging lifestyle environment. With this in mind, Gambrill Gardens is set to embark on a multi-million dollar renovation to set the standard for senior living communities in Ellisville.
The renovation of Gambrill Gardens will include a large communal area addition encompassing unique spaces that encourage socialization and interaction among the residents. This extensive remodeling beginning late 2016, will surpass $4 million when complete.
New and remodeled amenities will include enhanced lighting, a redesigned dining room, a casual bistro, an overnight suite for resident guests, fitness center, movie-room, salon, activity area, dining room and extensive outdoor patio areas.
The renovated and welcoming lobby and reception area has been designed to provide comfort, information, and easy access to the rest of the amenities. The corridors will be well-lit and hung with artwork and interesting wall coverings.
Enhancing the physical, social and emotional health of our residents is a primary goal in establishing a healthy senior community. The remodel will include an in-house gym/physical therapy room, Café, Arts & Crafts Room, Activity Room with sunroom where seniors can play games, mingle and bond with fellow residents.
Dining at Gambrill Gardens will be a nutritious and exciting experience in the new dining room and casual bistro. The kitchen staff will source food from the finest local vendors and fresh seasonal and locally available produce will be used everyday.


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Featured Links
Experience Soccer Travel to the Best Youth Soccer Camps in Europe, Register Online Now!
Coerver Coaching's ULTIMATE Bundle - Get Every DVD & Online Version
Benny Feilhaber scored the game winner in the final of the Gold Cup--a phenomenal volley that silenced the pro Mexican crowd in a match that was back and forth, with equally near goals for both Mexico and the US. Although Mexico was the more dangerous and skillful side in the first half, the US goal to tie it, a penalty kick, knocked the wind out of their sails a bit and the momentum swung to the US. Late in the game Tim Howard made a wonderful point blank save that enabled the US to hold on.
Read more...
Video by way of That's On Point
Quote of the Month:
"There is always someone out there getting better than you by training harder than you."
more soccer quotes...
© 2019 Soccer Training Info All rights reserved.


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The Effects of Brexit can be felt even before we leave, 10% hike straight off the bat on most things, Who are these idiots that compile RPI & CPI Stats, obviously they LIE. Grrr Filament Doubled in Price, if people only realized Stop Buying things & watch the Price Fall, Take your Money out of the Banks, and watch the Interest Rates Rise to Lure you Back! Take Control Build your Own Stuff!
After testing some cheap Black PLA, I think it might be time to find something better, So the search is on for some nice Matte Black Material. Or spend time on some finishing work.


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The Texas Dragon Boat Association (TDBA) is pleased to announce the Honorary Co-Chairs of the 9th Annual Gulf Coast International Dragon Boat Regatta taking place on October 20-21, 2012. The distinguished Honorary Co-Chairs include: The Honorable Vincent M. Morales Jr., Mayor, City of Rosenberg; The Honorable Li Guixi, Deputy Consul General of the People's Republic of China in Houston; Hillary Goldstein, Board President, First Colony Community Association; and Ronald A. DePinho, M.D., President, The University of Texas MD Anderson Cancer Center.
Teams from California, Louisiana, Arkansas, and Texas – including corporate, community, youth and breast cancer survivor (BCS) teams – will paddle 40-foot long dragon boats weighing 500 pounds in the waters of Brooks Lake in Sugar Land, Texas. "Dragon boating is a great wellness activity with cultural significance and has been embraced by breast cancer survivors all across the United States", said Dr. DePinho. "MD Anderson is pleased to support the Regatta."
The Regatta highlights the traditions and cultures of Asia and Asian-Americans. "Dragon boats represent an important cultural tradition in China dating to 278 B.C., and I commend the Texas Dragon Boat Association for sharing this tradition in the United States," said Deputy Consul General Li.


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The count down is on for the biggest birthday celebration in Nigeria this month..
Big Brother Africa favourite Tayo Faniran can't wait to hit the third floor come Friday 28th March 2016.
The Nigerian born South African based model and triple ambassador will be making 30 years in a few and he decided to liken his upcoming birthday to some characters in the Bible.
"Joseph became a prime minister at age 30, David became a king at age 30. As am clocking 30 this week, my level is changing for better in Jesus name. Amen," he stressed.
Meanwhile, the celebrations which will be in Nigeria have already seen the likes of Esther, Frankie board to Naija and so in simple terms, they are already in the hood in time for the birthday bash.
Happy birthday in advance Mr.Faniran.
For all the latest updates on Big Brother Africa / Big Brother Africa Season 10 Auditions, you can subscribe to receive email updates from www.BigBrotherAfricatv.com.
Like us on FACEBOOK: Facebook.com/BigBroAfricaFans & Follow us on Twitter @BigBro_Africa for more Big Brother Africa News and Updates


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Local artisans and craft-enthusiasts weren't the only ones to benefit from Etsy Vancouver Market: Made in Canada Marketplace this September. The third-annual Etsy Vancouver event raised $14,333 CAD for The KidSafe Project Society (KidSafe), a Vancouver-based charity with a mission to provide nurturing safe havens for vulnerable inner-city children when schools are traditionally closed.
Etsy Vancouver hosted a successful marketplace late September of this year, with more than 7,000 people in attendance during the two-day event. Half of each attendee ticket went towards KidSafe.
Confirmed by Danielle Rana, KidSafe Manager of Development & Community Engagement, the funds raised by Etsy Vancouver are equivalent to 35 children's spaces in the charity's 2-week Winter Break program.
We're so grateful to have community partners like Etsy Vancouver who can make our programs possible for our children!Danielle RanaKidSafe Manager of Development & Community Engagement
More than thrilled with the news, the Etsy Vancouver team is, once again, partnering with KidSafe for their upcoming event, Etsy Vancouver Market: Winter 2016. The event will take place on December 3 – 4 to celebrate the local Etsy community. Expect to see a variety of handcrafted goods such as handmade jewelry, home décor and skincare products.


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Tasia Nikiforos
By Merridy Granger
Year 6
Ascot Park
R-7 Gymnastics Focus School
Researched by students of
Ascot Park School
Supervising teachers:
Jennifer Harris
Dimitri Katsambis
Tasia Nikiforos was born in 1944 in a village called Pantansa which was very dry. She was one of eight kids living in a two storey house. She went to school when she was about six years old. She didnt have much variety of clothing to wear and worked to help out the family. She also grew her own vegetables.
Young Tasia went to school when she was about six years old. It would start at 8:00 every morning and she would come home each day at 12:00 to have lunch.
Tasia moved to Australia in 1964 when she was eighteen years old. Then after a year her brother came over by plane. Tasia settled in fine and she liked Australia a lot.
Then Tasia married Pandelis Nikiforos had two children of their own, Vikki and Alex.
Nowadays Tassia and Pandeli go to a club with lots of their friends and the club is called the Laconia club.


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My friend told me about this and it sounds like a great way to have guilt free sex ; I am pretty down to earth but have a wild side as well. I enjoy cooking,reading good books ,playing cards, going out from time to time and meeting people over the internet. I love to party at night and dance in the clubs.
I'll always try something once and if i like it i'll do it again! Comfortable in sweater and jeans or satin and lace. I am Long dark blonde hair hasel eyes.Girls like girls in maryland. I'm new to tumblr and I'm looking to make some new friends. So follow me and I'll follow back and we can be friends:). Graciouslady in Mount savage MD maryland - Naked girls: Corriganville MD maryland there i'm a good looking in shape girls for sex clean discreet. Watch Gaithersburg Maryland Girls Sex porn videos for free, here on ohtsevilla.com Discover the growing collection of high quality Most Relevant XXX movies and clips.
Have fun till then. Seeking 4 The Only 1 for Me I am not one to put my business out for all to see. © Copyright 2018


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... although still more than 6 hours to go of '09, here, in Chicago. Where I have already discovered (in Devon Market; all credits to Daniel) an interesting range of rather decent Romanian wines (eventually I have settled for a Feteasca Regala from Jidvei and, something special for tonight-- I am very curious!-- a Dracula's Blood Merlot). Good to know, given that I'll soon spend quite some time around here.
Anyway, let us enter 2010 on Massenet's Méditation de Thaïs, wonderfully interpreted by the one and only Leila Josefowicz (how can one not be in love with this woman?!). THE music piece for the New Year (why not)?


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Southwest Park Trip Part 1: Vegas to Zion2:10 PM
Last fall we hit the road for the most amazing vacation through some of the Southwest's best national parks. We've been lucky enough to have vacationed in Spain, Paris, Costa Rica... my husband and I both agree that our trip through Arizona and Utah was the best of anything we've done so far! You can check the full itinerary here, but we're getting you started at the very beginning in today's post -- flying into Las Vegas.
Why We Chose Vegas Honestly, it was an easy choice. We could get direct flights from NC to Vegas (direct flights are really rare for us), and both flight and car rental prices for Las Vegas tends to be cheaper than other nearby airports. Plus, knowing we were "roughing it" for the next week made me want to at least start out the trip in a crush of people, glitz, and good food. We rented an SUV for the trip because we knew we'd be logging a lot of car time, some of which would be on less than ideal road conditions. This especially came in handy driving around Monument Valley and in Arches National Park.
After picking up our rental car, we took a little drive down the strip like normal tourists and then set upon a Target to stock up on supplies. I'd recommend investing in:
- bottled water (we got two 24 packs and that made us through the entire week)
- first aid supplies like bandaids, neosporin, hand sanitizer (unless you already packed those)
- sunscreen
- nonperishable food/snacks: we got 2 tubs of trail mix, a few boxes of granola bars, beef jerky (you can't have too much), a bag of fuji apples, and a box of cereal (we are big time cereal munchers)
- a bottle of wine and a bottle opener if you always forget yours (don't judge)
- anything you forgot to pack --socks, sunglasses, toothbrush, batteries...
After loading up the car we made it priority number one to eat an obscene amount of food before making the 2.5 hour drive to Zion National Park. There were so many choices, but because we have watched an embarrassing number of Diners, Drive-Ins and Dives we had already staked out our destination months in advance: the Asian-fusion paradise called Bachi Burger.
|The oxtail chili cheese fries were a meal in their own. And worth every calorie.
|My man loves a good burger.
|So no surprise, but this was hands down the best meal of our trip.
Uncomfortably stuffed and with half of my burger relegated to a to-go box (to be happily consumed by Mark in a few hours -- no food ever goes long uneaten in my house), we headed out towards Zion National Park.
Where We Stayed
The Driftwood Lodge was a fantastic place to stay, located just outside of Zion's main entrance in Springdale, Utah. A free shuttle system runs through Springdale, bringing hotel guests to and from the park from morning til night. The shuttle pick up is just right outside of the hotel and couldn't have been more convenient. Our rooms were huge -- like I felt guilty with having so much space but maybe I had just gotten too accustomed to European hotel standards. We even had a little porch outside with a bistro table and chairs, perfect for uncorking a bottle of wine and enjoying the view. And the view is spectacular. The hotel backs up to a lovely mountain view with a little pasture where a neighbor's horses lazily grazed. Another perk? Their complimentary breakfast buffet was first class. Fueling up on bacon and coffee every morning with that kind of view was just a little peak at what we'd have in store for us at Zion.
|Pull up a seat; this was the breathtaking view from our room at the Driftwood Lodge
About Zion
Hands down, Zion National Park should be the top destination on your itinerary list. You could easily spend a week just exploring this park. The convenience is unparalleled here, as you'll soon realize when you start hitting up the other parks on this itinerary. There are well-maintained bathrooms (with actual toilets), picnic areas, well-marked trails, and insanely easy access with their fantastic shuttle service. There are hikes for all ages, and we spent almost our entire visit to Zion National Park with people in their late 50s or older. And believe me, it was inspiring because some of these older ladies were totally kicking my butt when we were hiking up Angel's Landing. #NoExcuses
Cost: $25 for a 7 day pass
And the views. It's unlike anything you've ever seen.
|The Patriarchs
Because we were only there for two full days, we had to pick just two major hikes to go on. I'd definitely recommend trying to get a third day if your trip allows because we had to leave out the Observation Point hike and I've heard great things about this one too. Just a reason to come back when I'm a ridiculously in shape 65 year old.
Top Hikes:
Angel's Landing -- heart thumping excitement; perfect to challenge your fear of heights
The Narrows -- do you like hiking in waist deep water?
Observation Point -- we didn't do this one but you get great views of the park and a view of Angel's Landing down below. Takes about 6 hours
Shorter Hikes:
Emerald Pools -- Not my fave but let me explain. There had been very little rain when we were there (well until the day we hiked the Narrows at least, but that's another story), so the "pools" were really just little mud puddles. Not scenic and crawling with toddlers and young kids splashing in the mud, running and nearly toppling the more elderly visitors that had chosen Emerald Pools because it's one of the easier hikes in the valley. I heard one older woman catching her breath on a rock whispering to her husband as she shook her head in dismay at the havoc of kids let loose like it was a mud and rock version of Chuck E Cheese "Their parents should have just taken them to the beach." Okay, not all trips are for ages 4-6 and maybe hiking the most popular elderly person trail in a national park is one of them. But when I have kids I'm sure I'll feel differently.
Weeping Rock -- If you're hot its nice to get a splash of water here as you walk "under" a little waterfall of rivulets.
Riverside Walk -- This is the start to the Narrows Hike, but even you don't plan on doing the Narrows, it's a nice little hike on a paved path that runs you right by the Virgin River.
I'd also really recommend just driving through Zion and stopping when you find something you want to explore. The changes in landscape are unbelievable. We got out and jumped around on the sides of mountains that looked like sandcastles and got to share the road with a herd of bighorn sheep. Not a bad way to spend an evening.
Want more Zion? We're talking Angel's Landing and The Narrows next!


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Iron and Manganese Removal
Iron, manganese and hydrogen sulfide can pose aesthetic problems for consumers and system performance problems for process water users.
Most groundwater contains some iron and manganese which naturally leaches from rocks and soils. Excess amounts in drinking water can cause discoloration, rusty-brown stains or black specks on fixtures and laundry, as well as affect the taste of beverages and can build up deposits in pipes, heaters or pressure tanks.
Conventional removal of iron, manganese and hydrogen sulfide has typically been accomplished by oxidation followed by filtration through a sand/anthracite media or by a combination of medias, some of which have the ability to accelerate the oxidation of the contaminants.
LayneOx enhances the kinetics of the removal process by serving as a catalyst in the presence of a pre-oxidant such as chlorine to effect contaminant reductions to well below the secondary MCL utilizing surface adsorption of the primary removal mechanism.