text:
Although iWeb can create Web pages you can upload to your own server, iPhoto has a built-in way of doing this as well that's simple and effective, even if its results aren't as visually interesting.
To export photos to Web pages:
Select an album or the individual photos you wish to publish via the Web.
Choose Export from the File menu (). iPhoto shows the Export Photos dialog.
If it's not selected, click the Web Page tab (Figure 5.40).
Enter the title for your Web page.
Enter the desired number of columns and rows of photo thumbnails.
If desired, select a background color or image.
Select the maximum width and height for the thumbnails and the full-size images.
Select the Show Title and Show Comment checkboxes as desired.
Click Export, navigate to the desired destination folder (it's best to create a new folder inside your user directory's Sites folder), and click OK. iPhoto exports the photos and builds the appropriate HTML files, naming everything for the enclosing folder.
Switch to the Finder, open the folder in which you saved your Web page, and double-click the HTML file to open it in your Web browser (Figure 5.41).
If necessary, upload the folder to your Web site using an FTP program like Interarchy (www.interarchy.com).
To manage plug-ins, select iPhoto in the Finder, choose Get Info () from the File menu, and use the checkboxes and buttons in the Plug-ins pane to enable, disable, install, and uninstall the plug-ins.
For a listing of plug-ins, check out the iPhoto Plugins News blog at http://wheezersociety.blogs.com/iphoto_plugins/.


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subset: commoncrawl


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wedding bands white gold - Ring For Women 2.3Ct Natural Aquamarine Ring Wedding Ring
Wedding Bands - A Symbol Of Never-Ending Love
The institution of marriage has undergone many alterations in excess of the many years, but the exchange of rings continues to remain an critical marital rite. The wedding ceremony is incomplete with no it as individuals thin, seamless bands worn on the ring finger on the left hand are not mere pieces of jewelry; they are replete with symbolism. They stand for enduring and immortal really like.
You will wear your wedding ring for the rest of your daily life. Consequently, choose an ornament that not only displays your marital dedication but also your individual style and taste. Here are ideas to select a set of timeless wedding rings for the bride and the groom.
Metal Choice
The 3 most widely used valuable metals are yellow gold, platinum and white gold. Gold has often been the traditional selection, but its yellow undertone can make it a little tough when it comes to coordinating it with clothes. White gold and platinum, with their silvery-white look are a greater selection in this regard. White gold is a more affordable option to the pricey platinum. When you purchase metal bands, make confident you examine the jewelry marking.
If you are seeking at less conventional alternatives, Rose gold seems lovely as well. It has a pink shade and is made by alloying yellow gold with copper. Tungsten bands are a appropriate choice for guys. Silver can also be utilised as a metal for wedding ceremony rings. Nevertheless, it is not the perfect materials because unlike gold, it is really soft and is simply broken, disfigured and scratched.
A wedding ceremony band is typically a seamless ring with out any gemstones and designs, but there is not harm in including one particular. A gemstone generates a dainty and delicate design and style. If you want to include a gemstone, consider of how you will location it. Also, if the bride's ring has a stone, it is not necessary for the grooms ring to sport a single; it looks greater plain.
Speaking of attainable gemstone selections, white diamonds are by far the most selection. Amethyst, turquoise, pearls and chocolate diamonds are other favorites for a wedding ceremony ring. When you get a ring with a stone, decide on 1 that has specific which means in your existence. It could be a birth stone. Decide on one that is difficult adequate to withstand every day dress in.
The fashion also adds to the beauty of the ring. There is a assortment of styles to select from. You can decide on a vintage fashion ring that expresses your cultural heritage or chunky, modern day ring with intricate patterns. If you've always loved to stand out in the crowd, opt for an unusual design wedding ceremony band. It could be a Goth band, horseshoe styled bands, dolphin rings and dragon bands.
Gold Wedding Bandsfor Men, White Gold WeddingRings, Gold Men'sWedding Bands, PlatinumWedding Bands, DiamondWedding Bands


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The Spaniard returned to the Blues starting line-up last night for their 1-0 Europa League win against Greece side PAOK after being left out again at the weekend.
The former Real Madrid striker was left frustrated though after missing several decent chances and was taken off with 10 minutes to go.
Morata shrugged off an attempted embrace from his manager and trudged towards the away bench, after another wasteful night in front of goal.
In the Premier League, Sarri has preferred to play Olivier Giroud in recent matches and the French World Cup winner has impressed.
And former Napoli boss Sarri believes that Morata is currently suffering a crisis of confidence.
The Italian tactician told reporters after the game with PAOK: "Alvaro has to gain confidence with one, two, three goals. I am not able to give him confidence.
"In this match he has had three, four opportunities. He was unlucky.
Despite failing to find the back of the net, the 59-year-old was pleased with his alert positioning.
"I have to say also, for maybe the first time in this season Alvaro was ready in the box. He was on the ball and was active," he added.
"So I am really very happy with the three points, very happy with the performance I think, but I am not very happy with the result. When it is time to kill the match we have to kill the match."
Morata – who could cost Chelsea up to £70m if all the add-ons in his deal are met – failed to fully impress last season.


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2014 64 VAUXHALL CORSA 1.2 S AC 5d 83 BHP
- 81816 Miles
- Petrol
- Manual
- 129 CO² (g/km)
- £125.00 12m Tax
- £68.75 6m Tax
- 51.4 Avg. MPG
- Group 6 Ins.
- 81816 Miles
- Petrol
- Manual
- 129 CO² (g/km)
- £125.00 12m Tax
- £68.75 6m Tax
- 51.4 Avg. MPG
- Group 6 Ins.
Vehicle Information
- Features
- Description
- Finance
- Gallery
Vehicle Summary
- Body styleHatchback
- Engine size1229cc
- Engine capacity1.2 L
- Fuel typePetrol
- Number of doors5
- Number of seats5
- TransmissionManual
- CO2 emissions129 g/km
- Insurance group6
Performance & Economy
- Fuel consumption (urban)39.2 mpg
- Fuel consumption (extra urban)62.8 mpg
- Fuel consumption (combined)51.4 mpg
- 0 - 60 mph13.9 seconds
- Top speed107.0 mph
- Cylinders4
- Engine power83 bhp
- Engine torque85 lbs/ft
- Height1488mm
- Height inclusive of roof railsmm
- Length3999mm
- Wheelbase2511mm
- Width1737mm
- Width including mirrorsmm
Interior / Exterior
- 4 speakers
- CD30 stereo radio/CD/MP3 player + aux-in socket
- Air conditioning
- Front seat back map pockets
- Reflection cloth upholstery
- Body coloured bumpers
- Dark style headlights
- Daytime running lights
- Door to door illumination
- Electric front windows/one touch/auto reverse
- Electrically adjustable door mirrors
- Follow me home headlights
- Rear wiper
- Matt chrome effect centre console
- 14" steel wheels
ComfortNo features available.
Safety / Other
- Service interval indicator
- Speed sensitive power steering
- Dual stage Driver/Passenger Airbags
- Electronic stability control
- Front passenger airbag deactivation
- Hill start assist
- Three rear inertia reel lap/diagonal seatbelts
- Tyre pressure monitoring system
- Immobiliser
- Remote central deadlocking
OtherNo features available.
Factory Fitted Extras
For more info on this vehicle call our showroom on 020 8589 9090
Every effort has been made to ensure the accuracy of the above information but errors may occur. Please check with a salesperson.
- Registered: 2014
- Mileage: 81816 miles
- Engine Size: 1.2 L
- Fuel Type: PetrolGearbox: ManualBody Style: HatchbackColour: SilverOwners: 2
For more info on this vehicle call our showroom on 020 8589 9090
Included in the price you will get a 3 months warranty with parts and labour. There are extendable warranty packages available at very low costs, which we can discuss with yourself on site. We are also proud to be a part of the AA dealer promise program which gives you 12 months breakdown cover with the AA at no extra cost. We work with a variety of different finance lenders to ensure you get the best deal possible, so contact us for any finance enquiries. We have over 45 cars in stock at any given time all available to view at www.roystoncars.com. If you have any questions regarding this car or any of our services please call us on 0208 589 9090 or 07940 031 001. Please feel free to pop in and browse through our stock at our forecourt in Hayes at 232 Yeading Lane, UB4 9AU. T&Cs apply. E&OE
Every effort has been made to ensure the accuracy of the above information but errors may occur. Please check with a salesperson.
Representative APR shown in example above.
Fees are already accounted for within the payments displayed and are also included within the total amount payable.
Finance available subject to status. Indemnities may be required. Other finance offers may be available but cannot be used in conjunction with this offer. We work with a number of carefully selected credit providers who may be able to offer you finance for your purchase. We are only able to offer finance products from these providers. Postal Address: 232 Yeading Lane, , Hayes, Middlesex, UB4 9AU
The following documents are available for this vehicle:


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September 15-October 15 is Latinx Heritage Month. We're inviting you to join us in reading stories by Latinx authors–and if you don't know where to start, here are a few of our recommendations!
We Are Not From Here by Jenny Torres Sanchez
A poignant novel of desperation, escape, and survival across the U.S.-Mexico border, inspired by current events.
Pulga has his dreams.
Chico has his grief.
Pequeña has her pride.
And these three teens have one another. But none of them have illusions about the town they've grown up in and the dangers that surround them. Even with the love of family, threats lurk around every corner. And when those threats become all too real, the trio knows they have no choice but to run: from their country, from their families, from their beloved home.
Crossing from Guatemala through Mexico, they follow the route of La Bestia, the perilous train system that might deliver them to a better life–if they are lucky enough to survive the journey. With nothing but the bags on their backs and desperation drumming through their hearts, Pulga, Chico, and Pequeña know there is no turning back, despite the unknown that awaits them. And the darkness that seems to follow wherever they go.
In this striking portrait of lives torn apart, the plight of migrants at the U.S. southern border is brought to light through poignant, vivid storytelling. An epic journey of danger, resilience, heartache, and hope.
Juliet Takes a Breath by Gabby Rivera
The Grief Keeper by Alexandra Villasante
Undead Girl Gang by Lily Anderson
The Fall of Innocence by Jenny Torres Sanchez
The Lovely Bones meets Celeste Ng for teens in this gorgeous, haunting, and tragic novel that examines the crippling–and far-reaching–effects of one person's trauma on her family, her community, and herself.
For the past eight years, sixteen-year-old Emilia DeJesus has done her best to move on from the traumatic attack she suffered in the woods behind her elementary school. She's forced down the memories–the feeling of the twigs cracking beneath her, choking on her own blood, unable to scream. Most of all, she's tried to forget about Jeremy Lance, the boy responsible, the boy who caused her such pain. Emilia believes that the crows who watched over her that day, who helped her survive, are still on her side, encouraging her to live fully. And with the love and support of her mother, brother, and her caring boyfriend, Emilia is doing just that.
But when a startling discovery about her attacker's identity comes to light, and the memories of that day break through the mental box in which she'd shut them away, Emilia is forced to confront her new reality and make sense of shifting truths about her past, her family, and herself.
A compulsively-readable tragedy that reminds us of the fragility of human nature.
Beyond the Break by Heather Buchta
Imaginary Borders by Xiuhtezcatl Martinez
Sanctuary by Paola Mendoza and Abby Sher
When I Am Through With You by Stephanie Kuehn
A gripping story of survival and the razor's-edge difference between perfect cruelty and perfect love.
"This isn't meant to be a confession. Not in any spiritual sense of the word. Yes, I'm in jail at the moment. I imagine I'll be here for a long time, considering. But I'm not writing this down for absolution and I'm not seeking forgiveness, not even from myself. Because I'm not sorry for what I did to Rose. I'm just not. Not for any of it."
Ben Gibson is many things, but he's not sorry and he's not a liar. He will tell you exactly about what happened on what started as a simple school camping trip in the mountains. About who lived and who died. About who killed and who had the best of intentions. But he's going to tell you in his own time. Because after what happened on that mountain, time is the one thing he has plenty of.


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It was not any type of Jacksonville FL amateur sluts local, no, not, but rather a arcing go throughout her lovely lap. He desired to create no fluster. Very slowly did he do it. He opened the car door by Jacksonville Florida statistic on casual sex across her, as noted lap. STANDING HER GROUND It had been associated by Cheery that once the door was open, he removed his shoes very carefully and slowly so as nobody with any urgency that was untoward. He turned to her from where he sat, with the door swung slowly open.
There are clues that they utilize that you can look out for. These include utilizing the expression" God Fearing" and saying" Am" rather than" I'm. " They'll write that age and space does not matter too. The guys might look much younger in their photos than they say they're around the profile.
TAKING COMMUNICATION OFF- LINE If you are able to trade replies and you'd like the individual at the other end, the goal should be to eventually fulfill; nevertheless, before you local sluts in person, you may choose to take the dialogue off the relationship website so that you local sluts who like to fuck Jacksonville talk with the individual in greater depth than a relationship newjersey sex dating sites permits.
I emphasize all of the Jacksonville FL that an individual will not sabotage himself from being his charismatic self in the interactions that he cares most about. We've always been, " on, " socially, in our own lives, and we have all faltered under stress- - I teach you to be the former all the time, particularly when it matters. Follow this advice, and magically, people begin reacting to you differently.
Here is one of the more benign responses among our female profiles using an Asian preference received from a Caucasian male: ( NOTE: We've blacked out any personal information like images, names, and phone numbers to local sluts the innocent and guilty alike. ) Cheeky, correct? She found what she was searching for, but if she examined her data than that which it required to bring out the worst in most folks she detected an even more interesting trend.
A girl in this way, will offer herself upfront to someone like me, who she's seemingly really attracted to or simply thinks is a high worth man( I swear, I am not trying to brag, I would like you to have these offers also) . She could very well enter a relationship with somestereotypical'niceguy' and make him wait for sex and never actually be that into it with him. But he is a fantastic provider and treats her nicely. Thus, she and he will date.
The Relationship Guru Rules I'm expecting you to do a great deal of dating over so I'm going to put some ground rules to be certain that you frustrated, overwhelmed or don't get exhausted. You aren't likely to fancy every single person so don't waste your valuable time on the people that are wrong you meet. Our mutual popular london dating apps Jacksonville Florida is to find Jacksonville FL local latina teen sluts of the partner of your dreams, although they may seem a little brutal.
Popular Online Internet dating Activities All types of daily activities are expanding in popularity online these days like sharing photos and also greeting cards. As a matter of fact, both of these even go together with on the internet meet local sluts Jacksonville chances, one of one of the most preferred online tasks for singles today.
Being casual about the company has huge benefits. Wheneveryou're a man that is casual, you come across as somebody who's enjoying his entire life and doesn't care about what others think or say about him- a caliber most women admire. If you come across as somebody who is heavily interested in relationship, you might do an overkill and turn off.
" No. Sorry. We finish our drinks and he says that while I despise cats and dogs, he does not care for your Normanby. And he is hungry. We should go someplace else.
However it looks like there's a greener lawn next door, so they hop once again! They do their companion jumping, exchanging and dating in search of the greenest grass, yet they'll never ever discover it due to the fact that a beautiful partnership, like a lovely yard, should be had a tendency to and cared for. You can have' gladly foreverafter' with the companion you local pregnant sluts Jacksonville FL, yet you have to devote yourself to it.
New condoms in your handbag. Do not depend on the man although most will, to have a source. If you bring your own, you will know about the freshness and the quality. Do not let him talk you.
Some type of relationship, call it chemistry. They were significant. I needed a reason to ask this individual on a date. More often than not, the telephone call did go farther than the relationship.
BIRTHDAY MESSAGE STARTER This is easy- - it is all about needing a birthday which catches her attention and makes her grin. Sadly, her boyfriend or man may take her out that day, so that day, that she may not respond or answer.
BUILDING A STRONG FOUNDATION While difficult conversations can range from embarrassing to frightening, they're necessary in developing a solid foundation. Just remember that you don't have to respond at the moment if your partner springs something. One of my favorite answers is, " You know, that is a fantastic point.
This could be with platonic friends or earlier spouses( see bulleted examples listed above) . Another situation could arise where one spouse has the chance to meet up with a former lover to local singles sluts Jacksonville FL java. Or perhaps, one partner beginning to fantasize about someone and is currently undergoing feelings she perhaps met on the Jacksonville FL local snapchat user names sluts, was introduced, or works with. Process her fascination with her partner and the spouse does not necessarily wish to act on those feelings, but wishes to share the expertise publicly. As you style your relationship built on trust, vulnerability, and 24, this instrument can be incredibly useful for local sluts no sign up or fres Jacksonville Florida your communication.
Remember? They're totally going to have your photograph and profile introduced to them as a new member, and you will be bombarded with flirts, winks, messages etc. .
The fuck buddy myvidster of empathy the covert narcissist has will make them not able to view you as a person. They will do whatever they need to, maintain and to gain control and power over you and also the relationship.
I've included a bunch of my discussions, as I've mentioned. There is much of a group arrangement. Meaning, it's not paint numbers. My face to face strategy with women, is a complete ceres male prostitutes, as I discussed in my book. I believe it is better to learn to read and respond on the fly as opposed to memorizing canned lines, although online dating has more structure.
Shortly after this I ended up in a nation in hospital and fell sick with a severe allergic response. I burned and came out of hospital one week after, alive, but still quite sick. Throughout my stay in hospital my buddy had decided that I wasn't worth the trouble and she ended it personally. I was in physical and psychological agony. I had that horrible, gut- wrenching pain of heartbreak.
Find a group of friends or a buddy with whom you feel secure- - friends you can trust. Describe this exercise and let them know that you are going to attempt and share a few of your masks together. By revealing your mask, explain that, it will not have the exact same power on you as it did before. Share with these friends from being honest and open and intimate with other people some of the fears that have prevented you. Ask the individuals to do the identical thing. Meaningful communication between you and your friends, open will allow you to get free and will help you recover some.
Because we are all human and no one is perfect, proclaim your innocence or examine them incredulously and inform them that they are being crazy and the response to reactivity is to get very defensive. This is not going to diffuse the situation. Here are a few hints that are little. Rule number one is that if somebody has been reactive and they say that they will need to really go for a walk, let them go for a walk. Jacksonville local hot and wet sluts is a reason for this. Rule number 2is to Jacksonville FL largest online dating services interested and curious as you can. You're going to bring in a few of being the observer, of those skills and you are likely to approach this with attention and curiosity. Though, I want to explain why you should let somebody go for a walk if they state they want to.
I had been mad and hurt- - hurt by his actions but angry in my stupidity. I had set myself. I was angry at having misjudged him. As his actions had revealed, was he cruel? Or was he simply unthinking? Or was he so emotionally detached that he could not think about anybody else but himself? He had been the first guy in the guys I'd dated that myself had opened up to- - my heart, my bed and my house. I had introduced my sister and him. We'd spent hours talking about things that mattered to all us to each other, sharing our previous experiences including a few painful and private happenings in each of our lives. Until then I'd found it hard to completely trust and open myself up to anyone, but I had done it with him, and I felt like this had been thrown back into my face. I felt overcome with the sense of loss.
Always listen. Never belittle or downplay what they imply or what they say. Let them explain their feelings or ideas in more detail so that they can be understood. Always let them know that you are there for them.
You'll discover that it does not matter what you say they'll be able to put a spin or a twist on it, so it means that which they need it to mean rather than what you meant. As narcissists are in love with themselves, they are also in love with using words against people. Their language that is wealthy draws people in, and farther down the street, it will be used to tear those very same people apart.
I am not here to judge you or to let you know what you should or should not put up in relationships or with in lifestyle. I'm only here to assist you help you align yourself to receive it and have a look at what it is you want.
Other questions might be, " How do I know when this is going to for you" " What does this look like? " " What can I do to you? " " What would you want from me whenyou're going through something like this? " He might actually say, " I need some space, " and I think that's a reasonable request if someone is requesting it and tell you that is what they desire at the moment.


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50 pubs listed for Thetford over 5 categories
Bars in Thetford
42 Annesdale, Ely Cambridgeshire, Ely, CB7 4BN
80 Main Street, Witchford, Ely, CB6 2HQ
19 Bridge Road, Mepal, Ely, CB6 2AR
8 Duck Lane, Haddenham, Ely, CB6 3UE
Burystead Bury Lane, Ely, CB6 2BD
128 High Street, Sutton, Ely, CB6 2NW
13 Victoria Street, Littleport, Ely, CB6 1LU
27 Admirals Way, Thetford, IP24 2LB
7 Market Place, Thetford, IP24 2AL
10 Anna Gurney Close, Thetford, IP24 1TA
45 King Street, Thetford, IP24 2AU
45 King Street Memorial Green Thetford Rd, Barnham, IP24 2PA
38 Norwich Road, Thetford, IP24 2JE
64 Magdalen Street, Coney Weston, Thetford, IP24 2BP
64 Magdalen Street King Street, Thetford, IP24 2AZ
5 Market Place, Thetford, IP24 2AJ
45 King Street Elveden Forest, Brandon, IP27 0YZ
64 Magdalen Street Elveden Forest, Brandon, IP27 0YZ
2 Anna Gurney Close, Larling, Thetford, IP24 1TA
The Green, Oxborough, PE33 9PS
7 White Hart Street, Foulden, Thetford, IP26 5AW
The Saxon Lynn Rd, Weeting, Brandon, IP27 0QW
1 High Street, Thetford, IP26 4NT
133 High Street, Lakenheath, IP27 9ET
2 Mill Road Elveden Forest Elveden Forest, Lakenheath, Brandon, IP27 0YZ
6 Angel Lane, Bury Saint Edmunds, IP33 1RF
Thetford Rd Thetford Road The Green, Thetford, IP24 3RU
Thetford Rd Thetford Road The Green, Thetford, IP24 3QU
Thetford Rd Thetford Road The Green, Thetford, IP24 2PU
Sports Bars in Thetford
27 Whitehart Street, Thetford, IP24 1AA
Watton Road, Thetford, IP24 2AD
Elveden Forest, Brandon, IP27 0YZ
Elveden Forest Elveden Forest, Brandon, IP27 0YZ
Elveden Forest Elveden Forest Elveden Forest Elveden Forest, Brandon, IP27 0YZ
Night Clubs in Thetford
Riverside Walk, Thetford, IP24 2BB
Riverside Walk, Stow Bedon, Thetford, IP24 1LD
Gastropubs in Thetford
Griston Road, Norfolk, IP24 3PG
Other Pubs in Thetford
1 Sandhill, Littleport, CB6 1NT
106 Main St, Little Downham, Ely, CB6 2SX
35 Old Market St, Thetford, IP24 2EQ
41 St Nicholas St, Thetford, IP24 1BE
Castle St, Thetford, IP24 2DP
85 Brandon Rd, Elveden, Thetford, IP24 3TP
7 Silver St, Witcham, Ely, CB6 2LF
45 High St, Wilburton, Ely, CB6 3RA
Caston Rd, Griston, Thetford, IP25 6QD
30 High St, Northwold, Thetford, IP26 5LA
69 Crown St, Brandon, IP27 0JU
Watton Rd, Wretham, Thetford, IP24 1QS
93 Castle Street, Thetford, IP24 2DN
Top bars in Thetford
7 White Hart Street, Foulden, Thetford, IP26 5AW
12 miles away
Now open again and run by 2 lovely people Aaron and Angie.
4 Market Place, Mildenhall, Bury Saint Edmunds, IP28 7EF
11.3 miles away
An absolutely fantastic place. Run by caring people who produce wonderful food and excellent service for their customers.
27 Admirals Way, Thetford, IP24 2LB
0.8 miles away
great food,always buy from here
16 The St Market Pl The Green High Street, Hopton, Diss, IP22 2AG
12.6 miles away
Found this by chance as we were staying up the road at the white house b&b . Lovely food and ...
Memorial Green, Larling, Norwich, NR17 1LE
12.6 miles away
The Pub is reopening this weekend for food. Thank goodness our village has got its pub back. Great new people ...
Top sports bars in Thetford
Sports Bars yet to be reviewed
Top night clubs in Thetford
Night Clubs yet to be reviewed
Top gastropubs in Thetford
40 The Street, Beck Row, Bury Saint Edmunds, IP28 8AE
11.6 miles away
Wonderful atmosphere and great food can be found here I well reccomend booking though if you want to eat
Norton Road, Bury Saint Edmunds, IP31 3QH
11.3 miles away
Don't bother if car park empty says it all! Frozen food bought in and warmed up as told by ...
Top other pubs in Thetford
Bell Ln, Barton Mills, Bury St Edmunds, IP28 6AJ
11.1 miles away
Great little local social pub in Barton Mills
43 Long Ln, Feltwell, Thetford, IP26 4BJ
11.1 miles away
the pub is under new management now. the new couple are friendly and welcoming. bar menu was good value. will ...
35 Old Market St, Thetford, IP24 2EQ
0.2 miles away
went for lunch on tuesday it was wonderful the food was home made presented perfectly tasted 1st class the dolphin ...
Twenty Churchwardens, Cockley Cley, Swaffham, PE37 8AN
14 miles away
Twenty Churchwardens is probably my favorite pub as it has the look, feel and food of a very traditional pub. ...
Forward Green, Stowmarket, IP14 5HN
19.9 miles away
Three words: excellent quality food


text:
Product Overview
Main Features
- internal
- 2.5"
- PCI Express 3.1 x4 (NVMe)
- Packed with a deep feature set, this cloud-inspired Intel 3D NAND SSD is optimized for cloud infrastructures, offering outstanding quality, reliability, advanced manageability and serviceability to minimize service disruptions
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text:
Welcome to WordPress. This is your first post. Edit or delete it, then start writing!
Govt jobs 2020 – Latest Government jobs | Naa Jobs India
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Human RNA helicase A was recently identified to be a shuttle protein which interacts with the constitutive transport element (CTE) of type D retroviruses. Here we show that a domain of 110 amino acids at the carboxyl terminus of helicase A is both necessary and sufficient for nuclear localization as well as rapid nuclear export of glutathioneS-transferase fusion proteins. The import and export activities of this domain overlap but are separable by point mutations. This bidirectional nuclear transport domain (NTD) has no obvious sequence homology to previously identified nuclear import or export signals. However, the Ran-dependent nuclear import of NTD was efficiently competed by excess amounts of the nuclear localization signal (NLS) peptide from simian virus 40 large T antigen, suggesting that import is mediated by the classical NLS pathway. The nuclear export pathway accessed by NTD is insensitive to leptomycin B and thus is distinct from the leucine-rich nuclear export signal pathway mediated by CRM1.
Macromolecular trafficking across the eukaryotic nuclear envelope occurs through the nuclear pore complexes and involves specific signal-receptor interactions. Nuclear localization signals (NLS) are capable of conferring the nuclear import and retention functions to heterologous nonnuclear proteins (3). Both the classical NLS, first identified in simian virus 40 (SV40) large T antigen, and the bipartite NLS, found in a number of Xenopus nuclear proteins, contain stretches of basic amino acid residues. NLS-bearing proteins interact with the NLS receptor, namely, importin α, in the cytoplasm. Subsequent binding to importin β targets the NLS-importin α/β complex to the nuclear pore for translocation across the membrane (12, 31). Another type of nuclear import signal, termed M9, was found in heterogeneous nuclear ribonucleoprotein (hnRNP) A1 and several related hnRNPs (26, 39). M9 specifies both nuclear import and export of hnRNP A1 and bears no sequence resemblance to the classical or bipartite NLS. The only known receptor protein for M9 is a protein distantly related to importin β, named transportin (1, 9, 36). Additional nuclear import sequences have been identified, but their receptors are currently unknown (16, 19, 27). A signal from hnRNP K is believed to bypass the requirement of a soluble receptor and interact directly with the nuclear pore (27).
Nuclear export of RNA and protein is less well understood but also appears to be signal mediated and energy dependent. The best-characterized, leucine-rich nuclear export signals (NES) use CRM1, also a distant relative of importin β, as their functional export receptor (6, 10, 32, 40, 43). Leucine-rich NES complexes with CRM1 and RanGTP in the nucleus prior to translocation through the nuclear pore. This complex formation is sensitive to leptomycin B (LMB). Different classes of cellular RNA were shown to use distinct pathways for nuclear export (18). Recently, the receptor for nuclear export of tRNA, named exportin-t, was shown to be another member of the importin β family (21).
The nuclear export of cellular mRNA is tightly coupled to splicing, such that only completely spliced mRNA is exported into the cytoplasm. However, export of unspliced viral RNA is necessary for the expression and replication of retroviruses. For lentiviruses, a viral regulatory protein named Rev promotes the nuclear export of unspliced and incompletely spliced viral RNA by binding to its cognate RNA sequence, the Rev response element (RRE) (4, 5, 24). Rev contains a leucine-rich NES and uses CRM1 as the export receptor (4, 6). In simple retroviruses, the unspliced RNAs contain acis-acting, constitutive transport element (CTE) that interacts directly with cellular factors to achieve nuclear export (2, 34, 41, 48). We previously showed that human RNA helicase A specifically binds functional but not mutant CTE in vitro (42) and that antibodies to helicase A blocked CTE function when microinjected into human cells (23). We also showed that helicase A shuttles constantly between the nucleus and cytoplasm despite its apparent steady-state nuclear localization (42). In this paper, we report the identification and characterization of a nuclear transport domain (NTD) in helicase A which directs the bidirectional trafficking of fusion proteins. This domain presumably plays an important role in the posttranscriptional regulation of retroviruses by helicase A.
Plasmid constructions.The cDNA clone of human RNA helicase A was a kind gift from J. Hurwitz (22). Plasmid pSK-helicase A was first digested with BglII, blunt ended with Klenow polymerase, and then digested with EcoRI. A 3.4-kb fragment that encodes amino acids 1 to 1136 of helicase A was recovered. pEGFP-1 (Clontech) was digested with BamHI, blunt ended, and then digested with NotI. A 0.7-kb fragment encoding the green fluorescent protein (GFP) was recovered. The 3.4- and 0.7-kb fragments were then used in a three-way ligation with theEcoRI-NotI-digested pcDNA3. The appropriate construct encodes amino acids 1 to 1136 of helicase A with GFP fused to the carboxyl terminus and is called GFP/HelA-ΔCTD. The GFP coding sequence from pEGFP-1 was PCR amplified and cloned into theBamHI and EcoRI site of pcDNA3 (Invitrogen) such that the EcoRI site at the 3′ end of the coding sequence can be used for in-frame cloning of GFP fusion proteins. This plasmid was designated pc-GFP. pSK-helicase A was digested with ApaI andEcoRI and then blunt ended with Klenow polymerase, and a 3.9-kb blunt fragment (including the 3′ untranslated region) encoding amino acids 137 to 1269 of helicase A was recovered and cloned into theEcoRI-digested but blunt-ended pc-GFP vector to make GFP/HelA-ApaI. GFP/ApaI-ΔCTD was made in a similar way, but theApaI-BglII fragment of helicase A was cloned into pc-GFP. To make the GFP-helicase carboxyl-terminal domain fusion proteins, different carboxyl-terminal domain fragments of helicase A were PCR amplified and cloned into the EcoRI site of pc-GFP. Site-directed mutagenesis was carried out by a method similar to that used with the Transformer kit of Clontech. Mutated sequences were used for testing subcellular distributions of the protein domain they encode. Myc-PK plasmid was described by Siomi and Dreyfuss (39). Wild-type and mutant NTDs were PCR amplified asKpnI-NotI fragments and cloned into theKpnI and NotI sites of pcDNA3/Myc-PK to make Myc-PK-NTD and Myc-PK-NTD mutant plasmids. Myc-NPc-TNLS has been described by Michael et al. (26). Wild-type and mutant NTDs were PCR amplified as XhoI-XbaI fragments and cloned into the corresponding sites of pcDNA3/Myc-NPc-TNLS to make NPc-TNLS-NTD, NPc-TNLS-ΔK62, and NPc-TNLS-ΔR65. GFP-TNLS-NTD and mutant constructs were made by cloning theEcoRI-XbaI fragments of the corresponding NPc-TNLS plasmids into EcoRI-XbaI-digested pc-GFP. pGST-NTD was made by cloning the EcoRI fragment of GFP-NTD into yeast vector pGAD10, resulting in pGAD10-NTD. TheBamHI-BglII insert of this plasmid was then cloned into the BamHI site of pGEX-2T (Pharmacia) to yield pGST-NTD. pGST-ΔK62 was made in a similar fashion.
Cell cultures, transfections, and Western blotting.HeLa and Cos-1 cells were maintained in Dulbecco's modified Eagle's medium with 10% fetal calf serum. Transfections were done by calcium phosphate precipitation. In experiments where transcription and translation inhibitors were included, they were added 45 h after transfection, 3 h prior to fixation of the cell for immunofluorescence assay. For the Western blot, Myc-PK-NTD plasmids were transfected into Cos-1 cells, and cells were harvested 48 h later and lysed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis buffer. DNA was sheared by passage through a syringe and boiling. Total proteins were either treated with alkaline phosphatase for 1 h at 37°C or directly loaded onto a 12.5% polyacrylamide gel. The separated proteins were transferred onto a nitrocellulose membrane and Myc-tagged proteins were detected with monoclonal antibody 9E10 (Babco).
Heterokaryon assay.Interspecies heterokaryons of HeLa and NIH 3T3 cells were formed as described previously (27). For GFP-CTD and GFP-NTD, cells were fixed 1 h after fusion. For the GFP-TNLS-NTD series, NPc-TNLS-NTD series, and Rev, cells were fixed 3 h after fusion. To confirm the inhibitory effect of cycloheximide on new protein synthesis, cells were metabolically labeled with [35S]methionine and [35S]cysteine 30 min after the addition of the translation inhibitor, labeled for 1 h at 37°C, lysed, and assayed for radiation incorporation.
Immunofluorescence analysis.For GFP-transfected cells, cells were fixed in 4% paraformaldhyde and viewed under a Nikon fluorescence microscope. For Myc-PK or Myc-NPc-TNLS fusion construct-transfected cells, the cells were fixed and stained with monoclonal antibody 9E10 as described previously (26). In all the heterokaryon experiments, 5 μg of Hoechst 33258 (Sigma) per ml was added either in 3% bovine serum albumin (BSA) after permeabilizing the cells with 0.2% Triton X-100 (for GFP fusion proteins) or during the secondary-antibody staining (for NPc-TNLS constructs). MEK-1 was stained with a polyclonal antibody purchased from Santa Cruz Biotechnology Inc. GST-NTD was stained with a monoclonal antibody against GST (Santa Cruz Biotechnology Inc.).
Recombinant protein purification, peptide conjugation, and microinjection.pGST-NTD was transformed into bacterial strain BL21. Recombinant protein purification was carried out with a commercial GST purification module (Pharmacia). The final protein sample was concentrated to 3 mg/ml for microinjection. NIH 3T3 cells growing on coverslips were injected by using an Eppendorf microinjector. For the LMB experiments, cells were treated with 20 nM LMB 6 h before injection. One group of coverslips was inverted onto 50% polyethylene glycol to form polykaryons; 1 h after cell fusion, GST-NTD was mixed with 1.5 mg of rhodamine-dextran per ml before being injected into selected nuclei of the polykaryon. The cells were fixed for GST staining 30 min after injection. Another group of coverslips was fixed for immunostaining with a polyclonal antibody against MEK-1. BSA conjugates were generated as previously described with either wild-type (CYTPPKKKRKLY) or mutant (CYTPPKTKLV) NLS peptide.
RNA gel shift assays.Gel mobility shift experiments were carried out as described previously (42). RNA helicase A protein and helicase A protein with the carboxyl terminus deleted were kind gifts from C. Lee and J. Hurwitz. GST-CTD was produced and purified similarly to GST-NTD.
The carboxyl terminus of helicase A harbors an NLS.N-terminal and C-terminal deletions of RNA helicase A were generated and fused to GFP (Fig. 1a). The subcellular localization of fusion proteins was compared with that of endogenous helicase A and of GFP alone. We observed that deletion of the N-terminal amino acids (amino acids 1 to 135) did not affect the nuclear localization of the fusion protein (GFP/ApaI). In contrast, deletion of the carboxyl-terminal domain (CTD) (amino acids 1137 to 1269) both in the full-length context (GFP/HelA-ΔC) and together with the N-terminal deletion (GFP/ApaI-ΔC) rendered the fusion protein completely cytoplasmic (Fig. 1b), indicating that the C terminus of helicase A is needed for its nuclear localization. To find out whether the CTD of helicase A is sufficient for nuclear import, we fused the CTD to GFP and examined the distribution of the fusion protein. GFP-CTD also had a nuclear localization similar to that of endogenous helicase A and GFP/ApaI. We therefore conclude that the CTD of helicase A is both necessary and sufficient for nuclear localization.
To map the minimal determinants for nuclear import, we fused a series of 5′ and 3′ deletions of the CTD to GFP and studied their subcellular localization. The minimal functional NLS was mapped between amino acids 1150 and 1259 (Table 1; Fig. 1b). The inability of further CTD deletions to localize in the nucleus was not due to their having insufficient sizes, because M9 of hnRNP A1, which is less than 40 amino acids long, was able to localize GFP in the nucleus in the same experiments (data not shown). Examination of this minimal NLS sequence revealed no homology to either the classical NLS (SV40 T antigen or bipartite NLS), M9 from hnRNP, or the nuclear import-export signal of hnRNP K (27, 39).
The CTD also contains a functional NES.We recently demonstrated that helicase A is a shuttle protein (42) and thus probably contains a nuclear export signal. We then tested the CTD for its ability to direct the nuclear export of GFP-CTD. Inhibition of transcription blocks the nuclear import of shuttling proteins, making it possible to detect these steady-state nuclear proteins in the cytoplasm (25, 35). In our previous experiments, actinomycin D treatment caused endogenous helicase A to accumulate in the cytoplasm of HeLa cells. Cytoplasmic accumulation of GFP-CTD was also observed in transfected cells similarly treated with actinomycin D (data not shown), suggesting that CTD is capable of mediating nuclear export.
To obtain more definitive evidence for the nuclear export function of CTD, interspecies heterokaryon experiments were carried out with GFP-CTD or the minimal NLS fused to GFP. HeLa cells that were transfected with these constructs were fused to NIH 3T3 cells to form heterokaryons. Cycloheximide was added to inhibit de novo protein synthesis. The appearance of GFP in the mouse nuclei within the heterokaryon indicated that the fusion protein had moved out of the human nuclei and into the mouse nuclei. The mouse nuclei were distinguished from the human nuclei by punctate staining with Hoechst 33258. A Myc-tagged form of NPc-TNLS (26), which contains the core domain of Xenopus nucleoplasmin and an NLS from SV40 T antigen, was cotransfected as a nonshuttling nuclear protein control. As shown in Fig. 2, GFP-CTD was exported from the nucleus of the transfected human cell while Myc-NPc-TNLS stayed in the same nucleus. Similar results were obtained for the minimal NLS domain (data not shown). Therefore, both CTD and the minimal NLS sequence also contain an NES. For this reason, we named the domain between residues 1150 and 1259 the NTD (nuclear transport domain). These results indicate that although GFP-NTD has a steady-state nuclear localization, it actually shuttles constantly between the nucleus and the cytoplasm.
The nuclear import and export activities of NTD are separable by point mutations.NTD, which contains part of the RGG box of helicase A (22, 46), is rich in glycine and serine residues. The N terminus of the domain contains several closely spaced basic residues (Fig. 3). Site-directed mutagenesis was used to identify specific amino acid residues that are critical for the import and export functions of NTD. Point mutations were introduced across the domain, and the mutants were tested for their ability to direct the GFP fusion protein in and out of the nucleus (Table 2). All the point mutants that retained the nuclear import function also maintained their nuclear export activity. Mutations in two basic residues, lysine 1162 and arginine 1165 (ΔK62, ΔR65, or R65L) abolished the nuclear localization of GFP-NTD, but deletion of arginine 1158 (ΔR58) or a neighboring tyrosine 1166 (ΔY66) did not have any effect. The loss of import function of the K62 and R65 mutants was confirmed in another experiment. NTD, ΔR58, ΔK62, ΔR65, and ΔY66 were fused to the carboxyl terminus of a Myc-tagged pyruvate kinase (39) and transiently expressed in HeLa cells. The subcellular localization of the fusion proteins was determined by immunostaining with a monoclonal antibody against Myc. Myc-PK localized exclusively in the cytoplasm, as previously reported (39). PK-NTD, PK-ΔR58, and PK-ΔY66 were all localized in the nucleus, while the PK-ΔK62 and PK-ΔR65 derivatives were localized in the cytoplasm (Fig.4a).
The efficient expression of the PK-NTD fusion proteins in mammalian cells was verified by Western blotting with the antibody against the Myc tag (Fig. 4b). PK was truncated at the carboxyl terminus when fused to the NTD series, and so Myc-PK-NTD is about the same size as Myc-PK. This analysis revealed that the PK-ΔK62 and PK-ΔR65 derivatives had a lower mobility than PK-NTD and PK-ΔR58. This reduced electrophoretic mobility was probably due to posttranslational modifications of NTD in the cytoplasm of eukaryotic cells, since GST fusion proteins of NTD and ΔK62 expressed in Escherichia coli exhibited similar electrophoretic mobility (data not shown). Treatment of the transfected cell lysates with alkaline phosphatase did not alter the mobility of PK-ΔK62 and PK-ΔR65 (Fig. 4b), suggesting that the shift is not due to phosphorylation.
ΔK62 and ΔR65 were further studied for their ability to mediate export. Wild-type NTD, ΔK62, and ΔR65 were fused to the carboxyl terminus of Myc-NPc-TNLS, and the resultant hybrid proteins were tested for their abilities to shuttle between the cytoplasm and nucleus by interspecies heterokaryon assays. As shown in Fig.5a, both mutants still shuttled. The ΔK62 and ΔR65 NTD mutations were also able to facilitate the shuttling of GFP-TNLS in heterokaryon assays (data not shown).
The effects of the ΔK62 NTD mutation on import and export function were further studied by microinjection analysis of proteins. A fragment encompassing ΔK62 NTD was fused to GST (GST-ΔK62). The expressed recombinant protein was purified and microinjected into either the cytoplasm or the nuclei of NIH 3T3 cells. When injected into the cytoplasm, the fusion protein stayed in the cytoplasm (Fig. 5b), but when injected into the nuclei, it became cytoplasmic after 30 min of incubation at 37°C (Fig. 5b). These results confirm the observations obtained in the transfection studies of the ΔK62 NTD mutation. Again, the ΔK62 mutation disrupted the nuclear import function of NTD while having no effect on its nuclear export. These results demonstrate that the import and export functions of the NTD are separable activities of the NTD.
Since the NTD contains part of the RGG box that is implicated in RNA binding, it was possible that the export of NTD was bridged by RNA (e.g., CTE) which was being exported by other NES-containing proteins. To test this possibility, the ability of GST-CTD to bind to the CTE was analyzed in a gel mobility shift assay. As shown in Fig.6a, although helicase A with CTD deleted had a weaker binding to CTE than did native helicase A, CTD itself was not sufficient for CTE binding in vitro. To further address the importance of cellular RNA in NTD export, RNase A was coinjected into the nucleus of Cos-1 cells with GST-ΔK62. The presence of RNase in the injected cells disrupted the integrity of the nucleus, as evidenced by the accumulation of the injection marker, rhodamine-dextran, in the nucleoli of these cells. However, GST-ΔK62 was exported efficiently in the same cells (Fig. 6b). Taken together, these results suggest that the export function of NTD is not mediated by binding to RNA.
The nuclear import of NTD can be competed with NLS peptides.Although the NTD sequence bears no resemblance to a classical or bipartite NLS, the fact that closely spaced basic residues are critical for import raised the possibility that NTD uses the importin α/β pathway. To address this question, excess unlabeled TNLS peptides conjugated to BSA were injected into the cytoplasm of NIH 3T3 cells along with both labeled BSA-NLS and GST-NTD. BSA conjugates containing mutant NLS peptides were used as a control for the specificity of the competition. Injection of fluorescein isothiocyanate-labeled BSA-NLS into the cytoplasm resulted in the nuclear import of the conjugates after 30 min of incubation at 37°C (Fig.7a), indicating that the BSA conjugates are functional as nuclear import substrates. In the presence of the BSA-mutant NLS conjugates, both the BSA-TNLS conjugates and GST-NTD were efficiently imported into the nuclei after 30 min (Fig. 7b, top panel). However, the nuclear import of both BSA-NLS and GST-NTD was competed by excess unlabeled BSA-NLS peptides (Fig. 7b, bottom panel). These results indicate that the nuclear import of NTD has the same sensitivity to excess NLS peptides as to conjugates containing TNLS. Microinjection experiments with T24N Ran also showed that the nuclear import of NTD is Ran dependent (data not shown), as expected for the classical NLS pathway. The glycine-rich feature of NTD also prompted us to determine if it interacts with transportin. NTD did not bind to transportin in the yeast two-hybrid assay, whereas the hnRNP A1 M9 region readily bound to transportin (data not shown).
The nuclear export of NTD is not sensitive to LMB treatment.We tested if the nuclear export of helicase A NTD is sensitive to LMB, which disrupts the function of the leucine-rich NES. GST-NTD was injected into selected nuclei of NIH 3T3 cell polykaryons in the presence of 20 nM LMB, a concentration which is 10 times that used to inhibit Rev export (45). Normal shuttling of GST-NTD would be evidenced by its appearance in the uninjected nuclei within the polykaryon (Fig. 8a). The efficacy of LMB treatment on nuclear export in this study was controlled by analysis of the mitogen-activated protein kinase kinase (MEK-1) in parallel. MEK-1 is a shuttle protein with a steady-state cytoplasmic localization and contains a leucine-rich NES at the amino terminus (11). While LMB treatment relocalized MEK-1 to the nucleus by blocking the NES-mediated export as previously reported, it had no effect on GST-NTD export (Fig. 8b). This result indicates that the export mediated by NTD functions in a CRM1-independent manner.
RNA helicase A contains a sequence that specifies bidirectional nuclear-cytoplasmic trafficking.ATP-dependent RNA helicase A was first purified from HeLa nuclear extracts as an in vitro RNA helicase activity (22). A subsequently described nuclear DNA helicase II (46) is the same protein. Helicase A contains two RNA-binding domains, a helicase core and an RGG box. Recent studies suggest that this protein may be involved in gene regulation at both the transcriptional and posttranscriptional levels (23, 29, 42). In spite of its predominantly nuclear localization (47), helicase A apparently shuttles between the nucleus and the cytoplasm (42). Here, we showed that a domain of 110 amino acids at the carboxyl terminus of helicase A is a bidirectional NTD. Helicase A with a deletion of this domain accumulated in the cytoplasm, in contrast to endogenous helicase A or GFP-helicase A fusion proteins that contain the NTD. NTD also targeted a completely cytoplasmic protein, pyruvate kinase, to the nucleus upon fusion to its carboxyl terminus. These observations suggest that NTD is both necessary and sufficient to mediate the nuclear localization of RNA helicase A. In addition, the NTD contains a nuclear export signal, since it was able to direct the rapid export of GST, GFP, a nuclear form of GFP (GFP-TNLS), and NPc-TNLS in a variety of assay systems. The import and export activities of NTD are separable by point mutations in two basic residues (K62 and R65), which abolished the ability of the NTD to direct nuclear import while sparing the export activity.
Nuclear import activity of the NTD.Although no appreciable sequence homology was found between NTD and other NLSs, a short stretch of amino acids relatively rich in basic residues is found in NTD, the U1A signal, KNS, and the NLS of Sam68. An arginine-to-alanine substitution was shown to abolish the NLS function of Sam68 (16). While both K1162 and R1165 are crucial for the nuclear import function of NTD, R1158 is dispensable for this function. Interestingly, PK-ΔK62 and PK-ΔR65 mutants had a lower gel mobility than the nuclear counterparts (PK-NTD, PK-ΔR58), which had the expected mobility. It is possible that NTD is modified at the posttranslational level in the cytoplasm and that this putative modification is related to its NLS activity. We showed that this modification was not at the level of phosphorylation. Despite the lack of sequence resemblance to classical NLS, NTD apparently uses the same import pathway, since its nuclear import can be efficiently competed with an excess of BSA-conjugated NLS peptides.
Nuclear export activity of NTD.Nuclear export activity of NTD was demonstrated in several different systems. First, NTD was able to direct the export of three different heterologous proteins in interspecies heterokaryon assays. GFP-NTD, GFP-TNLS-NTD, and NPc-TNLS-NTD all shuttled when tested, indicating the NTD can direct nuclear export even in the presence of a strong NLS. Second, export and reimport of GST-NTD were detected within 30 min after being injected into selected nuclei of NIH 3T3 polykaryons, demonstrating the rapid shuttling ability of NTD. Finally, a mutant form of NTD that lost its NLS activity directed nuclear export upon injection into the nucleus. This export function of NTD is not mediated by RNA binding. The NTD showed no ability to bind to the CTE in vitro, and coinjection of RNase had no effect on export function.
Helicase A and Rev use distinct nuclear export pathways.Leucine-rich NES have been found in a variety of cellular and viral proteins (4, 7, 8, 15, 28, 37, 44). The human immunodeficiency virus Rev activation domain is a prototypic NES that is involved in the nuclear export of intron-containing human immunodeficiency virus mRNAs. It has been shown recently that CRM1, a cellular protein distantly related to importin β, serves as the functional export receptor for Rev NES. Competition experiments withXenopus oocytes suggest that CTE and Rev-RRE utilize different cellular factors for RNA export (34, 38). Therefore, if helicase A plays a role in CTE-mediated export, it should function in a CRM1-independent manner. LMB disrupts the complex formation of NES, CRM1, and RanGTP by binding to CRM1, thus blocking Rev export. In functional studies, LMB blocked Rev-RRE-mediated but not CTE-mediated gene expression (33). We found that NTD-mediated export was resistant to 20 nM LMB, a concentration 10-fold higher than that previously observed to block Rev-dependent mRNA export (45), indicating that NTD does not use CRM1 as its export receptor. This observation is consistent with a possible role for helicase A in CTE-mediated export. Our recent observation that the injection of antisera specific for helicase A perturbs CTE function in human somatic cells further supports an important role of this protein in CTE function. The human protein TAP was recently also identified as a cellular CTE-binding protein that is involved in CTE-mediated export in Xenopus oocytes (13). The importance of TAP and that of helicase A in CTE function are not mutually exclusive. Rather, both proteins may be required to facilitate efficient RNA export. Alternatively, the two proteins may function at different steps in the CTE-mediated processing and export of unspliced RNA expressed by type D retroviruses. Further functional studies will help address the biological significance of helicase A in CTE-mediated RNA export.
Why simple and complex retroviruses have evolved different pathways to achieve the same function is not clear. By requiring the accumulation of an early viral protein, Rev, to direct the nuclear export of partially spliced and unspliced RNA, complex retroviruses are temporally regulated and may thus have an advantage in delaying immune system surveillance by the host and building up a larger burst size for viral production. Simple retroviruses are often less cytopathic and express lower levels of virus. For this type of chronic infection, a constitutive cellular export pathway seems to be sufficient. Although simple and complex retrovirus mRNAs apparently use distinct cellular receptors for nuclear export of unspliced mRNA, they may still have some common step(s) in their posttranscriptional regulation, e.g., in their interfacing with the splicing machinery to make unspliced RNA available for export. Our recent data (23) that helicase A may be involved in the Rev transactivation pathway at a step prior to nuclear export are consistent with such a hypothesis.
We thank C. G. Lee and J. Hurwitz for the helicase A cDNA, Gideon Dreyfuss for Myc-PK and Myc-NPc-TNLS plasmids, B. Wolff for LMB, M. Vodika and M. Emerman for NLS peptides, T. R. Reddy for help with the constructions of pGST-NTD, W. D. Xu for help with site-directed mutagenesis, K. Kuhen for critical reading of the manuscript, and F. Gage and the James B. Pendleton Trust for the use of their microscopic facilities. We also thank C. Goodwin for his excellent technical assistance throughout the study.
This study was supported in part by NIH grant AI35477 to T. Hope and NIH grant GM56089 to F. Wong-Staal.
- Received 31 August 1998.
- Returned for modification 5 October 1998.
- Accepted 29 January 1999.
- Copyright © 1999 American Society for Microbiology


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Why every senior caregiver should consider professional training
If you're a senior care provider, you help aging individuals with their activities of daily living. You keep them company during your shift, and you care for them like they're your own family. But to provide the best care and to fully connect with families you want to work with, it's important to invest in your caregiving skills.
Hear what other senior caregivers say ongoing education has done for them and how you can benefit from it in your career.
Education and training ensures quality of care
Without specialized training and guidance from care professionals, you may feel inexperienced and unsure of what's best for your charges. You may be trying your best, but lack of experience and knowledge can be a huge risk — to you and your charge.
"When I first started working with seniors, I felt like I didn't know enough to properly care for my charges," says Jerrica Allen, a registered nurse from Thornton, Colorado. "I also worried that my CNA certification didn't give me the skills I needed to really connect with them."
Professional elder care providers should be trained properly to provide the best quality of care possible. Continuing education, or "refresher" training, can remind you of best care practices. Frequent training also offers an opportunity to learn more about your newest senior charge, who will likely have different needs or conditions from your previous ones.
You need to be qualified to do the work
A senior care provider is not a medical professional. If you want to provide medical services to your charges, you will need the certifications to do so.
"Caregivers that provide any quality of living attention, such as giving showers, feeding and assisting in personal care, should be a certified nursing assistant," says Dr. Barb Norvell, a podiatrist and vice chair of Podiatry at WellStar Atlanta Medical Center. "More complex medical attention, including passing of medications and administering medications, should be performed by a certified nursing assistant."
It may actually be illegal in your state to provide medical services without the proper training, so make sure you are qualified and up to date on certifications.
Families trust your skills and experience
Families want to know you're prepared and trained for anything that may happen. They're putting their trust in you as a professional caregiver. Not having enough training or being underprepared is a big risk to your charge's health. Families may also be looking for specialized care — and your training can make you an easy hire.
For example, elder individuals are more susceptible to health conditions, such as Alzheimer's. Caring for someone with a condition like Alzheimer's may be beyond the scope of basic daily care some families can provide, and a care provider who has knowledge in this condition would be more appealing to that family as they seek to hire someone.
Knowledge prepares you for harder times
Part of providing care for seniors is, unfortunately, watching them age. If they have a severe medical diagnosis or are advanced in age, it can be very emotional for both you and the family. However, with the right training, you can actually be more equipped to help these families.
Sandy Griffin, LPN, hospice and palliative care nurse, and quality assurance coordinator at Hospice of South Louisiana, believes caregivers should study grief counseling and alternative care methods. Learning about bedside manner and how to communicate about death and illness can be a huge help, she says, as can discovering ways to distract your charges from pain or negative emotions.
"Art classes, book clubs and other social or private events to engage your charges will do wonders for them," Griffin says.
There are plenty of great books you can read on the subjects of aging and death, including "The Art of Aging," "Life's Final Season" and "Being Mortal." Finding a local hospice facility and asking if they have courses or resources for caregivers is another great way to get support and advice from people who are the real experts in "the hard stuff."
Education reinforces your desire to care for others
When a family hires an elder care professional to look after a loved one, they want to find someone who is properly trained and experienced — and who really cares. It's in your blood to care for others, and incorporating ongoing education and training into your work is the best way to do just that. At the most basic level, learning something new can reaffirm your desire to help others and may even lead you to your new family.


text: We booked online and got a great price. What a Surprise! Upon arrival, we found a beautiful clean lobby. Our room was more spacious than we had seen online for other more expensive hotels. The bed was super comfortable. With the price of our room, we received a hot breakfast buffet of scrambled and hard boiled eggs, potatoes, sausage, cereal, yogurt, bread, bagel. The room was big. A little dark in the bathroom and no mirror in the room but it was fine. The staff was kind. We will definitely remember this one for future stays.


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A mixed use, cash flowing opportunity! Located in a high visibility, high exposure commercial area - this fully occupied mixed use property boasts 3 updated one bedroom apartments with views of Beluga Lake (W/D in units!), and a 2,300sf office/retail commercial space below. Multi family tenants pay ALL utilities. Commercial tenants pay electric, gas & refuse.
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This contract for buyer services (the "Buyer Services Contract") between Boutiques Connection buyer (the "Buyer") and Boutiques Connection of Roseburg, OR ("Boutiques Connection," "our," or "us," and collectively, the "Parties") governs Buyer's access, use, and purchase of products and services on http://www.boutiquesconnection.com and http://www.boutiquesconnection.com domains including any content, functionality and services (the "Site"). By opening an account, Buyer agrees to be bound by the terms and conditions of this Buyer Services Contract, the Terms of Use, any documents the foregoing agreements expressly incorporate by reference, and any additional terms and conditions found on the Site (the "Agreements").
This Buyer Services Contract may be changed by Boutiques Connection without prior notice, at any time, in Boutiques Connection's sole discretion. The latest version of this Buyer Services Contract will be posted on the Site. It is Buyer's responsibility to check periodically for changes to the Buyer Services Contract before accessing, using, or purchasing any products or services through the Site. Your continued use of the Site after a posted change in the Buyer Services Contract will constitute your agreement to such changes.
Electronic Platform Provider
Buyer acknowledges and agrees that Boutiques Connection is an ecommerce platform provider that facilitates the marketing and sale of goods. Buyer further understands that Boutiques Connection is not a direct party to any sales transactions conducted on the Site, which are freely conducted between, and at the sole discretion of the Buyer and vendor. Accordingly, Buyer further understands and agrees that Boutiques Connection bears no responsibility or liability for claims of loss from transactions on the Site, or from conflicts and disputes between Buyer and vendor, or between the Buyers.
No Warranty
Buyer acknowledges and agrees that Boutiques Connection makes no guarantee or warranty with respect to any claims made by any vendor on Boutiques Connection regarding pricing, quality, origin, shipping cost, delivery date, availability, return/refund, suitability or authenticity of any items listed for sale on Boutiques Connection..
Information and product listings on Boutiques Connection may contain typographical errors, inaccuracies or omissions. Boutiques Connection therefore reserves the right, but not the obligation, to cancel transactions based on errors, to correct any errors, inaccuracies or omissions, and to change or update information at any time without prior notice.
Account Suspension
At its sole discretion, Boutiques Connection has the right to modify or terminate this Buyer Services Contract at any time for any reason, or for any action or behavior deemed as harmful or disruptive to other members (Buyers or vendors), or in any way detrimental to its business, reputation and image. Boutiques Connection reserves the right to rescind, suspend or terminate the Buyer's account at any time, without cause or advance notice, for any reason.
Site Issues
In the event of Site issues or malfunction, Boutiques Connection will undertake to work diligently to correct the problem in a timely manner. Boutiques Connection shall not be held liable for any claims of loss made by the Buyer as a result of Site maintenance, or as a result of events beyond the control of Boutiques Connection, including but not limited to, natural disasters, riots, wars, malicious software attacks, communications, power, and utility disruptions.
Boutiques Connection may assign this Buyer Services Contract at any time, in whole or in part, to any affiliate of or successor in interest to Boutiques Connection, without Buyer's consent. Buyer may not sell, assign, or otherwise transfer this Buyer Services Contract, and/or any rights or obligations or other interest under the Buyer Services Contract, in whole or in part, whether voluntarily, involuntarily or by operation of law including through any change of beneficial ownership that occurs in connection with a merger (including a reverse triangular merger), acquisition, or the sale or transfer of all, or any part of Buyer's business or assets (in each case, an "Assignment") without Boutiques Connection's prior written consent in each instance which consent may be granted or withheld within Boutiques Connection's sole and absolute discretion, and may require the acquirer to enter into an agreement with Boutiques Connection acknowledging and transferring obligations under the Buyer Services Contract to the acquirer. Subject at all times to the foregoing sentence, the rights, obligations and other interests of Boutiques Connection and Buyer under the Buyer Services Contract shall be binding upon and inure to the benefit of Boutiques Connection and Buyer, and to their respective successors, executors, administrators and permitted assigns. Any Assignment made in violation of this section shall be void.
The Agreements constitute and contain the entire agreement and understanding between the Parties with respect to the subject matter herein and supersede and replace all prior negotiations and proposed agreements, written or oral. The Parties acknowledge that no representations, inducements, promises, or agreements have been made by or on behalf of any Party, except those covenants and agreements embodied in the Agreements.
Governing Law and Jurisdiction
This Buyer Services Contract shall be governed by and construed in accordance with the laws of the United States and the State of California without regard to its conflict of law provisions. The Parties shall endeavor to resolve through good faith negotiations any dispute arising under or relating to the Agreements before proceeding to binding arbitration. If a dispute cannot be resolved through such negotiations within a reasonable time, either Party may request for claims to be resolved by binding arbitration to be conducted in Roseburg, Oregon by the American Arbitration Association ("AAA") under applicable AAA rules then-in-effect. Payment of all filing, administration and arbitrator fees will be governed by the AAA's rules, except as provided in this Section. To begin an arbitration proceeding, either Party must send the other Party written notice requesting arbitration and describing such Party's claims. Any and all proceedings to resolve claims will be conducted only on an individual basis and not in a class, consolidated or representative action. The Federal Arbitration Act and federal arbitration law apply to this Vendor Services Contract. An arbitration decision may be confirmed by any court with competent jurisdiction.
Any notice to Boutiques Connection must be in writing to the address set forth above and will be deemed given: (i) upon receipt if by personal delivery; (ii) upon receipt if sent by certified or registered U.S. Mail (return receipt requested); or (iii) one (1) day after it is sent if by next day delivery by a major commercial delivery service, with a copy via electronic mail.


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Say goodbye to your basic top! The Helena asymmetric Top is made from stretchy poly fabric, featuring an asymmetrical neckline, fitted bodice in babe loving crop length.
* 70% Cotton |30% Polyester* Asymmetric* Length 12" | Size S fits bust 32 - 33"* Model wears Size S
TWFS sizes are cut to fit the following body measurements. Do use it as guide for purchase.
(conversion metrics: 1" = 2.54cm)
How to Measure: To choose the correct size for you, measure your body as follows
When in doubt, wear demin. Shop now for the right fit.
Brand new look for 2020! Thank you for being with us
There is beauty in simplicity. Shop the minimalist collection


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wow. What a stupid thing to do. Risking his health and losing out on $24 million.
Sad, and I don't mean in an unhappy way.
BREAKING: New York Mets second baseman Robinson Cano has tested positive for a performance-enhancing drug and will be suspended for the entire 2021 season, sources familiar with the situation tell ESPN. He will forfeit a $24 million salary. News story will be up soon at ESPN.
10 Team, Points Only, Cash League
25 Man Roster (no position), top 20 point getters count at end of month
Keep 20/25 at seasons end, Cut 5 to FA for redrafting
Goalie points W=2pt L=-1pt SHO=2pt
Stamkos, Tavares, Eichel, Couture, Wheeler, Lee, Gusev, RThomas, Kucherov, Nugent-Hopkins, Burns, Zuccarello, Fabbri, JSchwartz, KPalmieri, KConnor, Necas, Yandle, Point, Konecny, TBertuzzi, NSchmaltz, Vrana
G- Vasilevskiy, Bobrovsky
"Cleavage is like the sun. You can look, but dont stare.. Unless you're wearing sunglasses."
wow. What a stupid thing to do. Risking his health and losing out on $24 million.
12 team, H2H, salary cap
1 pt/G or A, 2 pts/W, 1 pt/OTL, 1 pt/SO
Forward: B. Point, A. Svechnikov, K. Fiala, D. Larkin, R. Thomas, TBD
Defense: M. Giordano, A. Boqvist, TBD, TBD
Goalie: P. Grubauer, D. Kuemper
League has been running continuously since 1997-98
As if the first time wasn't enough
Yesterday's weirdness is tomorrow's reason why. - H.S.T.
I get when fringe players, AAA guys do steroids, a player going into his FA yr, but never stars with contracts. I just don't get it. Must be an ego thing.
What this does from a baseball aspect, it opens up $24mill the Mets were already planning on spending for FA. Might make them an option for Lindor now too
12 team, H2H, 6 keeper league (Keepers in Bold)
G, A, Pts, +/-, PPP, PIM, FW
W, GAA, S%, Saves


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The Property Manager spoke with Morin's this morning, they indicated they will start the final clean-up on Tuesday, February 10, 2015, starting at 9 am. They will sound their horns when they are in your area doing the final clean-up, so please move your cars.
With all the snow we have had in a short amount of time, it is increasingly getting harder to put snow in their normal locations when plowing. Morin's and Management is asking that when Morin's is the area, that ALL cars be out of the way. This means that no car should be in the way of the clean-up by either parking on the sides of the roads and leaving it there or not moving your car because you have shoveled around it. Your vehicle will be towed if you are impeding the snow removal process.
If you can't park your vehicle in your driveways or garage, please park at one of the clubhouses (located on Tinker Road, Glastonbury Drive and Kessler Farm Drive). Many villages have overflow parking as well.
Shoveling of decks should be done PRIOR to the final clean-up. If you shovel your deck after the clean-up is complete in your area, it will be your responsibility to remove the snow.
Walkers, dog walkers, and children should continue to be aware of the limited visibility due to the high snowbanks and make way for plows. You may be able to see and hear them, but they may not be able to see you.
Waste Management has informed the Property Manager that trash/recycle pick-up will delayed by one day this week. Trash/recycle will be picked up on Friday, February 13, 2015. DO NOT put your barrels and bins out before 8:00pm on Thursday, February 12, 2015.
If your unit is rented, it is the unit owner's responsibility to provide this information to the tenant. You can forward the email sent by the Property Manager or direct them to
http://www.kesslerfarms.com Latest News block/Twitter feed.
If you have any questions, please contact the Property Manager at firstname.lastname@example.com.


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SAN FRANCISCO (CBS SF) — A wet weather system spinning to life in the warm, humid air around the Hawaii Islands was taking aim at the Bay Area Monday, threatening to bring several inches of rain later this week, forecasters said.
The moisture-rich atmospheric river — called by some the 'Pineapple Express — was forming off the Islands and is more common in the winter months. On a scale from 1 (weak) to 5 (exceptional), the National Weather Service said the front will batter Northern California starting late Thursday for about 48 hours at category 3 or category 4.
The weather service tweeted that the storm could drop as much as 3 inches of rain on San Francisco by Saturday — which historically would rank the storm among the 10 wettest starts for April.
While the storm system will not impact the San Francisco Giants home opener on Tuesday it could create havoc with the schedule when the Los Angeles Dodgers roll into town for a four-game series on Thursday.
While the storm will be packing plenty of moisture, warm temperatures will drive the snow level above 8,000 feet in the Sierra. Most ski resorts will have rainshowers instead of snow flurries on the lower runs.
While 2 1/2-3 inches were forecasted for San Francisco more than 3 inches of rain was predicted from Ukiah to Monterey to Sacramento. The coastal mountains could see 4 inches or more.
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Answered By: Peter Nagels Last Updated: Jun 16, 2020 Views: 3
As the majority of our students are now completing their course work off campus, our library services are running on a limited basis.
Check out our opening hours below.
If you place an item on hold, you will only be able to pick it up from Broadmeadows Library as this is the only library which is currently staffed. If the item needs to come from another campus, there may be a slight delay.
Broadmeadows Library (for basic library services and computer access)
Monday - Thursday: 8:30am - 5:00pm
Friday: 8:30am - 4:00pm
Access to computers at the Broadmeadows Campus is also available in Building J, Room
207 from: 8:30am - 5:00pm
Docklands Library
Essendon Library (Unstaffed - computer access only)
Monday - Thursday: 8:00am - 8:30pm
Friday: 8:00 am - 6:00 pm
Richmond Library (The book collection is closed - computer access only)
Monday - Friday: 8:30am - 5:00pm
The e-library is open 24/7 https://libguides.kangan.edu.au
You can contact the library via webchat, or email us at email@example.com
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Contact Us
You may also contact us by using one of the methods below.
LLC phone number: 9279 2424
email: email@example.com


text: Home Stock Ant Group gets Chinese nod for Hong Kong leg of $35 billion... Stock Ant Group gets Chinese nod for Hong Kong leg of $35 billion dual listing: sources By - October 19, 2020 0 18 Facebook Twitter Pinterest WhatsApp RELATED ARTICLESMORE FROM AUTHOR Biden win lifts world stocks to record peak; dollar fades Wall Street set to jump after Biden election win Norwegian Air suffers body blow as government rules out more support LEAVE A REPLY Cancel reply Please enter your comment! Please enter your name here You have entered an incorrect email address! Please enter your email address here Save my name, email, and website in this browser for the next time I comment. Latest posts Could Brexit be the construction sector's greatest challenge? New COVID-19 cases below 1,000, lowest since Nov. 4 Gov't lifts total ban on conferences, workshops Transport dep't signs usufruct agreement for 5 DND properties for Metro Manila Subway project Anti-corruption task force approves guidelines to speed up complaints processing


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He was a first-time nonviolent possible offender, ... And under the mandatory minimums, he was put in prison for 15 years. Not only does the punishment not fit the crime, but the mandatory minimums don't give judges any discretion to look at the background of the case, to read into the specifics of the case. I don't know a judge who really is in favor of the mandatory minimums.
It is hard, but it is excellent, to find the right knowledge of when correction is necessary and when grace doth most avail.
Whatever you think of de Sade, he was a complex figure and we should not look for easy answers with him. He was, strangely perhaps, against the death penalty, and he was never put in prison for murders or anything like that.
We judge ourselves by what we feel capable of doing, while others judge us by what we have already done.
I never saw a man who looked With such a wistful eye Upon that little tent of blue Which prisoners call the sky.
Prosecution I have managed to avoid; but I have been arrested, charged in a police court, have refused to be bound over, and thereupon have been unconditionally released - to my great regret; for I have always wanted to know what going to prison was like.
Mere factual innocence is no reason not to carry out a death sentence properly reached.
If punishment reaches not the mind and makes not the will supple, it hardens the offender.
Virtue pardons the wicked, as the sandal-tree perfumes the axe which strikes it.
Prison, dungeons, blessed places where evil is impossible because they are the crossroads of all the evil in the world. One cannot commit evil in hell.
The law does not pretend to punish everything that is dishonest. That would seriously interfere with business.
The best situation of all, and one frequently utilized, is for jails and prisons to allow volunteer ministers of all faiths to enter prisons and offer their services to the inmates who want them. That way, the religious needs of inmates are met but without government funds being spent.
Fast closed with double grills And triple gates–the cell To wicked souls is hell; But to a mind that's innocent 'Tis only iron, wood and stone.
There is no peace because the making of peace is at least as costly as the making of war - at least as exigent, at least as disruptive, at least as liable to bring disgrace and prison and death in its wake.
No written law has been more binding than unwritten custom supported by popular opinion.
Three hundred years ago a prisoner condemned to the Tower of London carved on the wall of his cell this sentiment to keep up his spirits during his long imprisonment: "It is not adversity that kills, but the impatience with which we bear adversity."
When the Nazis came for the communists, I remained silent; I was not a communist. When they locked up the social democrats, I remained silent; I was not a social democrat. When they came for the trade unionists, I did not speak out; I was not a trade unionist. When they came for me, there was no one left to speak out.


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C, United States Army, jobs Chief Surgeon, American Expeditionary Forces. Semple died at Philadelphia on was graduated from the Medico-Chirurgi'cal College Dr. Before describing the method under consideration, I shall endeavor to mention a few of the points at fault in some wellknown methods. Mowat's cases of convulsions in children treated successfully by chloroform-vapour, it occurred to me others measures had been employed; but, finding the convulsions becoming almost continuous, I gave twenty minims of spirit of chloroform every hour, with the effect of soothing the patient, and in about twelve hours completely arresting the convulsions.
This is the veritable demand upon the doctor: titanmed.com. The condition of the patient, however, made it necessary to make the attempt, and accordingly I performed the operation this An incision was made through the integuments with some degree of obliquity, between three and four inches long, beginning a small distance above ihe tumour, and extending upwards externally to a middle line between the umbilicus and superior anterior spinous process of the ilium.
On the other hand, the waters of the New River, East London, and Kent pretty clear, it contained a high proportion of organic nitrogen", indicating very recent sewage contamination. In Guy's, his left hand became quiet, but not his right.
NOTES TOWARDS THE HISTORY OF THE MEDICAL STAFF OF THE ENGLISH ARMY PRIOR TO command of the army of Aulus Plautius, crossed the Thames, and took Camalodunum (Colchester) by siege.
But the care of the sick has little sanitary bearing. Canmed.com - the gastrointestinal symptoms seen in several patients after tho symptoms of severe shock had disappeared were reflex and dependent upon the laryngotrachcal-bronchial irritation In the fatal case the following gross appearances in the gastrointestinal MEDICAL ASPECTS OF GAS WARFARE The stomach contained a small amount of thin, grayish fluid, slightly blood stained, and some milk curds. Enjoyed of witnessing the employment of this plant in different diseases in the New-York Aims-House, and from the observations and experience of several distinguished practitioners, he was led to believe it an important and efficacious remedy in the treatment of most febrile disorders, in intermitting and remitting fevers; in yellow fever artd other diseases of specific contagion; in many cutaneous affections, and in diseases of general debility.
These reactions are likely triggered by the methylprednisolone cleanmed.com salts rather than the base corticosteroid. The best results are obtained by making simultaneous pressure through the rectum by means of a thick sound or bougie. The second was that of James Cole, who, while labouring under a delusion that his wife had hidden people under the floor and in the cupboard to try to poison him, took up his unoffending child of three and a half years from its cradle and killed it by knocking its head violently against the wall.
SOBOBON TO THK DISPENSARY OF ST. If I closed my left eye I could see very little indeed; and though the mistiness and movement were visible when the left eye was open, I was then able to see things much better than when it was shut. A survey of the list of exhibitions a.'id scholarships to which we have alluded shows that it is not one College alone, but many, which are thus desirous of encouraging a taste for natural science; and, when we consider that these numerous and valuable prizes will be awarded in a single year, wc must acknowledge that Cambridge is doing her part towards raising the.standard of medical education.
Mix the decoctions, and when clear, concentrate the liquor by evaporation.
In the epigastrium, slightly to the right of the median line below the liver margin, numerous hard nodular masses, slightly tender and movable, were made out.
Early in his illness he was very tolerant of the efTusion, and he would frequently go until a hundred ounces or more would be drawn from his chest, but later on in the disease his dyspnea would be extreme, and only thirty or forty ounces would be withdrawn, and toward the end he w-ould suflfer more dyspnea from a small quantity of fluid than he had formerly suffered when the large quantity These frequent operations were of necessity a great tax upon my time, but I found that by simplifying and systematizing the procedure I could sterilize the instruments, my hands, and the site of the puncture and complete the vapping in about twenty-five minutes, at the same time doing the operation aseptically. The size of the tube may vary according to the back water common to the mill. The deficient oxygenation gives rise to widespread temporary injury used in shells, and scattered from them on the ground, where it slowly evaporates. Reduce dried rhatany root to a moderately fine powder; mix it with half its weight of distilled percolation with temperate distilled water; Concentrate the liquid, filter before it becomes too thick, and evaporate it in a vapour-bath to a due consistence.
With these essentials at the clincian's command, the ultimate outcome will depend upon his readiness to employ them in time, and with such which brings us to the second factor essential to the prevention of terminal conditions. The absence of leucocytosis is consistent with the quiescent condition of the pocketed pus, there being no evidence of inflammatory defense. In odd moments he plays upon the mandolin, guitar, harp, ukulele, and the rest of the time on the ladies' heart strings. A more accurate way of estimating capacities of ourselves and other people is needed so as to place the right man in the right place.
Its use has the advantages of enabling one to know the intraabdominal pressure at any time, and is less tedious in use than a syringe: www.anmed.com.pl. Pillars approximated with four silk Omentum ligatured and ablated.


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Karen Moore, M.ED., B.ED. BA-Psych, RP Qualifying
Soul Path. Karen's passion is to ensure that people of all ages feel empowered to live their life to the fullest.Through individual sessions, classes and workshops, Karen will help you realize your gifts and potential! Relax & enjoy modalities such as Reiki or Reflexology or discover your soul's needs and gifts through Soul Talk self empowerment classes or Parent Groups on Resiliency Skills for those with Anxiety!
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Karen is a graduate of Spiritual Psychotherapy. Psychotherapy and Counselling is available for children, teens or adults. Karen spent 20+ years in the education system as Administrator, Teacher, Special Education/Resource Teacher and Behaviour Specialist. She is registered with the Ontario College of Teachers, and has a Guidance Counsellor designation. Karen uses a variety of modalities to address Life Transitions, Relationships, Grief, Depression, Anxiety, ADHD, Mental Health issues and assists helping professionals with compassion fatigue and burnout.
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A free twenty-minute phone consultation is available for those unsure of whether psychotherapy is the right approach for them.


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Inside the Spanish School Madre Tierra, Monte Verde Tours offers hikes, tours and bike trips around Xela. They organize day trip to Santa Anita and Nueva Alianza coffee farms.
13 Avenida 8-34
Quetzaltenango , Guatemala
Tour Operator Types:
Hiking/walking/trekking. wildlife tours.
Languages Spoken
Spanish, English
Established in:
More than 10 years
Currencies accepted
US dollar, Guatemalan quetzal
Contact Information:
Zip: Zona 1
Phone: (502)7761-6105
E-mail: info(at) monte-verdetours.com
URL: www.monte-verdetours.com
Enter your username and password here in order to log into the website:


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|Directory Services
The System container of a domain partition holds per-domain operational information. This includes the default local security policy, file link tracking, network meetings, and containers for Windows Sockets registration and resolution (RnR) and RPC name service (RpcNs) connection points. The system container is hidden, by default, and provides a convenient place for storing objects that are of interest to administrators, but not to end users.
Services that are not tied to a single host may want to create their SCPs under the System container of a domain partition. This alternative can be useful for services with replicas installed on multiple hosts, each replica providing identical services to clients throughout the domain. It enables you to group all the objects for the replicated service under a single container.
Services that create service-specific objects in the System container must do the following:
A vendor with multiple products can use a similar strategy to group service-related objects for all of its products. In this case, you could create a Container object with a name that clearly identifies the vendor; then create Container objects for each service as children of the vendor container. Create the vendor-specific container as a child of the System container.


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4 people prayed for this
You can putvyou trust in man and roll the dice or you can put you money on God and let God handle the man. When one is walking with God his path is straight and thus safe to follow.
I am placing everything I have on JESUS CHRIST OF NAZARETH.who has already won.