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Figure 4

-Deglycosylation of Epo-Epo fusion protein. Epo-Epo was treated with -glycanase, followed by SDS-PAGE and Western blotting (see “Experimental Procedures”). Note the narrow band of ∼42 kDa, consistent with full-deglycosylation. A minor band of ∼38–39 kDa is also visible and probably represents Epo-Epo lacking -linked sugars.

Epo-Epo is highly biologically active in vitro . We transfected COS1 cells with either conventional Epo wt cDNA or Epo-Epo (Epo wt /Epo wt ) cDNA. After 3 days, the supernatants were harvested and subjected to an in vitro bioassay. Protein was measured by ELISA calibrated against recombinant human Epo (Table bar strap sandals Grey Giuseppe Zanotti Cheapest 5V4PFKN8gM
). Epo wt was expressed at modest levels (3.5–9.2 IU/ml; 0.010–0.026 μg/ml) and exhibited a mean specific activity of 352 IU/μg (13 IU/pmol). This specific activity is slightly higher than that obtained by us previously for wild-type Epo expressed in COS1 cells (201–284 IU/μg) ( 10 ). In marked contrast, we found a much higher level of expression of Epo wt /Epo wt (110–195 IU/ml; 0.109–0.194 μg/ml), which exhibited a substantially higher mean specific activity (1007 IU/μg; 76 IU/pmol).

View this table:
Table I

Biological activities and protein concentrations of Epo and Epo-Epo fusion protein in transfected COS1 cell medium

Both Epo domains of Epo wt /Epo wt are biologically active. We showed this by mutation of arginine 103 (or the arginine 103 equivalent in Domain B, which is arginine 223 of the fusion protein) to alanine (R103A). This mutation results in complete inactivation of the Epo wt protein ( Outlet Find Great Black OverTheKnee Sock Boots Miu Miu New Fashion Style Of Sale 100% Guaranteed 2KAt5asH
, 20 ). We prepared three different constructs that incorporated this R103A mutation: ( a ) Epo R103A /Epo wt , ( b ) Epo wt /Epo R103A , and ( c ) Epo R103A /Epo R103A . We reasoned that if each of the two domains were active, then mutating only one domain would result in a protein that still retained biological activity, but with some decrease in specific activity. Only the double mutant Epo R103A /Epo R103A should be inactive. As seen in Table I , both Epo R103A /Epo wt and Epo wt /Epo R103A were expressed efficiently, and both exhibited biological activity. Interestingly, the mean specific activities of the two mutants were 480 (36 IU/pmol) and 516 IU/μg (39 IU/pmol), respectively, essentially one-half that of the non-mutated Epo wt /Epo wt (1007 IU/μg; 76 IU/pmol). This result strongly suggests that introduction of the R103A mutation into each Epo domain resulted in inactivation of that domain, allowing the other, wild-type domain to activate the receptor. This indicates that both Epo wt domains of Epo wt /Epo wt are biologically active, i.e. independently capable of receptor activation, and nearly equally so. The double mutant Epo R103A /Epo R103A was expressed at the mRNA level (data not shown). However, no activity or protein was detected in the COS1 supernatant.

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Posted on November 8, 2016 by Callan Bentley | Leave a comment

Notice of Meeting

Association of Environmental Engineering Geologists Date: Wednesday 11/16/2016 SECTION MEETING

Baltimore MD – Washington DC – Harrisburg PA

TOPIC: Geopier Rammed Aggregate Pier™ Applications Regional Case Studies

TOPIC: Geopier Rammed Aggregate Pier™ Applications Regional Case Studies

PRESENTER: Ms. Shana Carroll, P.E., Northeast Preconstruction Manager; GeoStructures, Inc.

PRESENTER: Shana Carroll, P.E., Northeast Preconstruction Manager; GeoStructures, Inc.

Rammed aggregate pier systems are a ground improvement technology that can be used to increase bearing capacity and control settlement of buildings, tanks, Retaining walls and embankments to close tolerances. Depending on site requirements, RAP systems can be installed using replacement (Geopier GP3®) or displacement (Geopier Impact®) methods. RAP systems are used to reinforce good to poor soils, including soft to stiff clay and silt; loose to dense sand; organic silt and peat; variable, uncontrolled fill; and soils below the ground water table. They can be used as an alternative to deep foundations or over-excavation, and provide cost and time savings on sites where poor soils and/or existing undocumented fill are a concern in terms of bearing capacity and settlement. This presentation reviews fundamental Geopier design and construction techniques and includes regional case studies such. Projects discussed include the National Cancer Institute’s Advanced Technology Research Facility located in Frederick Maryland and underlain by karst geology in the Piedmont Province, the Gaylord National River Ballroom located in National Harbor, MD in the Coastal Plain Province, and the George Mason University Fenwick Library in the Piedmont Province.


Shana Carroll currently serves as Northeast Preconstruction Manager for GeoStructures, Inc. and was formerly an Engineer with GeoSyntec, Inc. Shana has been involved in the design, estimating, and pre-construction management of specialty geotechnical systems throughout the East Coast since 2001. Shana holds a B.S. M.S. degree in Civil and Environmental Engineering from Bucknell University and is a registered Professional Engineer in 5 states. She is the immediate past president of the Baltimore Chapter of American Society of Professional Estimators (ASPE) and on the Program Committee for the National Capital Section Geotechnical Executive Committee of the American Society of Civil Engineers (ASCE). In her spare time, Shana enjoys writing about topics related to young professionals on her blog ( ) and traveling.


Electronic Filing

You must use the Iowa eFile system to file electronically unless you get permission from the court to file in paper. For instructions on using the Iowa eFile system, see the Sale Striped Glitter Espadrilles Anniel Anniel Great Deals Cheap Price Cheap Price Pre Order Real p1E4nxsLxC
and User Guide .

Conduct and Procedures that are Important to Know before Going to Court

Do not miss your court date . If you have a reason why you cannot go to court on the assigned day or at the assigned time, call the clerk of court’s office in your county. If you fail to appear on a scheduled court date for a civil action, the judge may enter a default judgment against you and find in favor of the other party.Also, the judge may find you in contempt of court. If you fail to appear on a scheduled court date and you are facing criminal charges, you may face additional criminal penalties, including jail, fines, and license suspension.

Be on time . If you can, visit the courthouse before the date of your hearing to see how people act, dress, when they arrive, and where to go. Arrive early to the courthouse on your court dat: plan fortraffic, parking, and nervousness.Watch forsigns and notices telling you where to go. In many court rooms, you should tell court staffthat you have arrived. If you are not in court when your case is called, and the other party is present, you might have a default judgment entered against you. If neither party is present when the case is called, your case may be postponed until the end of the day orfor another day or week.

Know and follow court rules .You shouldreview the applicable Iowa Court Rules . Judges will not make exceptions for self-represented litigants.

Be courteous and respectful .Be courteous and respectful to everyone: the judge, jury (if there is a jury), the other party, and attorneys (if they are represented).Wait for your turn to speak. Do not interrupt the judge or the other party. The judge wants to hear from each party and all partieswill be allowed to present their side of the case. You will be given the chance to respond to what the other party says, or you may politely ask to respond to the last comments.Do notinterrupt others in court: the judge will stop you and instruct you to wait your turn. Please turn your cell phone off or on silent.

Dress appropriately . The courtroom is a place of tradition, civility, and good manners. You should wear nice clothing: a dress, suit,sport coat or blazer and pantsas if you are going to church or a funeral.Do not weara hat or cap while you are in the courtroom or judge's chambers.

Be prepared . You may want to contact an attorney before deciding to represent yourself.You should understand the impact and consequences of the court action.Before your court date,determine whatthings you need to prove or defend your case. Bring anydocuments, photographs, receipts, witnesses, etc. that will help prove your claim or defend aclaim against you.

Even "small" cases can be legallycomplicated. If you do not understand something about your case, you should contact an attorney. Judges and court staff, including clerks of court, cannot give legal advice.

Speak loudly and clearly . Many people are nervous when they are in court and tend to speak softly. Judges, court reporters, and the other parties need to clearly hear what you say.

Frequently Asked Questions

Can judges and court staff assist with my case?

Court staff may explain basic procedures and answer questions about deadlines and fees. However, Iowa law and ethical rules prohibit judges and court staff from giving legal advice. This means that judges and staff cannot advise people on whether to bring claims to the court, what remedies they should seek, and the proper course of action.

Can I talk to the judge about my case without the other parties present?

No. Judicial ethics strictly prohibit judges from "ex parte" communication—which means talking with oneattorney or party without the otherattorney or party present.

If you want to give the judge informationabout yourcase or you want the judge to take some actionon yourcase, you must file your request with the court.

How do I find a lawyer?

Ask your friends, family, and colleagues for a referral or use the Iowa State Bar Association's Womens 26372 Trainers Marip Factory Price Outlet Store Locations Discount Recommend Outlet 2018 New Wholesale Price Cheap Price hOybeP1v
onlineor toll free: 1-800/532-1108.Court personnel are not permitted to make referrals to specific attorneys or law firms.

How do I request an ADA accommodation?

Please contact your local ADA coordinator to request an ADA accommodation. You can find your ADA coordinator by using the county directory or this Clearance Footaction Clearance Limited Edition Womens 42ma207700591 Trainers 8 Dockers by Gerli Cheap Low Shipping Discount Recommend M3xEw

Information on this web page is general in nature. Descriptions of laws and court procedures are abbreviated. This information is not intended as legal advice.If you do not understand this information or if you need legal advice you should seean attorney.


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