text:
We're into the third week now of our course 'Sarah and healing our self-esteem', and what a time it has been!
We've been working with Sarah's symbol, doing affirmations for self-esteem, chants for self-worth, prayers, and exercises for forgiveness. I've been working away at the programme along with the rest of the group, and Sarah has been overlighting all of our healing processes with her divine presence. So much has been, and is being 'lifted' from us in the group, and I feel I'm really experiencing divine grace in action…
Each week, I've been sending out Sarah's 'blessings' to the group (this is a way of transmitting Sarah's energy), and each time it's been completely different but had a definite theme each time; gentle grace, strength and this was one week's email out to the group:
"Dear all, last nights blessing had a very different feel about it. Last week was gentle and full of grace, this week it was focused and strong. There was such strength there, it was amazingly and awesomely strong!
To me it felt like energy to wake us up to just how powerful we are, I really felt that. (so the strength is what is inside us)
I can't say my mind has caught up with it yet, but I usually find with these things, that the energy comes first and then everything else gets dragged along with it.
So expect some changes this week! (the wind of change is in the air!!)
Of course I know with the blessings, that we all receive what we need…"
And it has been an honour to behold, the power and strength, grace and beauty coming through to each member of the group, as we all let go of what we no longer need, and find all of these wonderful qualities inside ourselves!!
Last week we did a forgiveness exercise, and after I had this email from a lady called Ginny in the group,
"Something quite magical happened to me after I'd finished the Forgiveness exercise on Saturday! The doorbell rang and there was my 90 year old neighbour standing on the doorstep, a lovely guy called Arthur. He bowed and then presented me with a beautiful light pink rose, just the colour of the heart chakra! He's never done anything like this before and the timing was just perfect! The rose was so beautiful I took a photo." (which is the photo at top of blog)
I loved the synchronicity of this, and I noticed straight away the three buds – Sarah often shows herself in 3's as she holds the energy of the threefoldflame; you can see a channelling which talks about this http://surrey.thespiritguides.co.uk/Article_Ascended_Master_Lady_Sarah__the_Age_of_Aquarius_594.aspx.
…So Sarah's energy has been overlighting our work, showing her presence in special ways.
And of course, working with self-esteem can mean some difficult things come up, but Sarah's energy seeps through everything, so her light and joy emerges in the darkest of times!!
This has shown itself in the 1:1 sessions each person has had with me and Sarah, and we have removed ancient blocks, healed our inner children, loved ourselves and brought back parts of our souls and re-membered our Divine Selves.
But no matter how difficult and dark some of the issues have been that have come up, the gentle love, the Divine grace, the brightness of ourSelves, the forgiveness in understanding; in the face of these gifts, the darkness becomes something to be held and loved once again, bearable in it's wholeness. Sarah holds the light and the dark within unity consciousness; you can see more about this http://surrey.thespiritguides.co.uk/Article_The_Ascended_Master_Sarah_who!_Why_Sarah_is_here_and_what_she_
So with each session comes a profound joy for me, as I experience Sarah; her love and beautiful light and her grounded and womanly wisdom.
She knows what it is to be human, and has a special joy and delight in helping us to empower ourselves.
Her whole presence empowers, and helps us to be more of who we are!
The learning for me is very deep too, as I see the strength, power and wisdom in others, so I see it in myself, and have realised that I have much to value in myself, even though I am currently living and moving amongst many that have masters degrees as a minimum qualification, and Phd's coming out of their ears (and here's me with my little diplomas in nursing and hypnotherapy!)
So this has been my biggest realisation on this self-esteem course so far, that like Sarah, it too is my joy to be able to facilitate greater self-esteem in others, and bring about greater self-empowerment.
That the gifts I have been given to tune into another's awareness and energy body, and sense there the issues, that need healing brought to them, and the ability I have to hold Sarah's energy around us, as we complete this process; that now is the time for me to honour and appreciate them – as all of the group are finding what is special within them, finding their strength, so I am too.
We are finding ourSelves!
And this is perhaps where our self-esteem comes from I think, seeing what is already there, seeing the truth, seeing our own beauty.
Seeing ourSelves.
And that was the message for me, in the rose above, and that's what I wrote to Ginny,
"and Sarah is on all sides, (in the 3 buds) and I just want to point out one more time, that that beautiful rose in the centre
If you would like a one-to-one session with myself and Sarah working on your self-esteen, I'm offering a 10% discount until the end of October, to book, you can find my contact details http://www.sarayei.com/contact-us/.


text:
Homeowners across the nation are discovering that they can save on energy costs when they specify Structure, Quest and Alside's Prodigy® Vinyl Siding instead of ordinary vinyl siding. To calculate your potential energy savings, go to www.ProdigySiding.com and use the online Energy Savings Calculator.
"Quality, Service and Selection from a Company You Can Trust!"
Maintaining the curb appeal of your painted home can be very costly and time consuming over the years.
A decision to invest in vinyl siding provides the immediate benefits such as:
- A virtually maintenance free exterior
- Superior energy cost savings and energy efficiency
- Increased property value
- Coordinated trim and Accent Options
- Clear-Through Color…never needs sanding, painting or staining
Prodigy® and Structure is designed with an R-Value of 5.0.
That's five times the energy efficiency of ordinary siding.
From its flawless appearance, unsurpassed insulation, maintenance freedom and
Limited Lifetime Warranty (including an industry leading fade warranty), Prodigy is the embodiment of what you value.

subset: commoncrawl


text: The King Bayside Spa Suites offers the comfort of our guest rooms, but with an additional parlor area that allows for that extra space some guests may need. The parlor area features a sleeper sofa, and is separated from the bedroom with French doors. These suites are located in the Spa Building with a bay view. It is the perfect option for a small family traveling on vacation, or for a romantic get-away.


text:
It has been incredible to see the response of our customers as we have helped them to understand and acquire this tax credit. With only a few months remaining, we continue to see potential home buyers coming in and wanting to be a part of our program, the
Salt Lake City, UT (PRWEB) July 7, 2009
Holmes Homes, an award-winning new Utah home builder dedicated to providing high quality and energy efficient homes in Utah, Arizona, and Idaho, announces the "Federal Tax Credit Advantage". Holmes Homes wants to help potential home buyers take advantage of $8,000 federal tax credit for the "first time homeowner". In an effort to stimulate the economy and revive the housing market, Congress has enacted legislation providing a tax credit of up to $8,000 for all first-time home buyers.
Time is of the essence for buyers who want to take advantage of this opportunity. Only homes purchased on or after January 1, 2009 and before December 1, 2009 are eligible. Don't forget that the definition of a "first time homeowner" is anyone who hasn't owned a home in the last 3 years, or that has never owned a home. This means all renters, recent college grads, and fence sitters need to get moving and come see Holmes Homes.
"There is no question that the combination of historically low interest rates and the $8,000 federal tax credit has dramatically increased interest in our homes, which has translated into increased sales over the last few months. We have been fortunate to have this wonderful tool to aid and assist our homebuyers; it has been great!" exclaimed Ross Holliday, Partner in Holmes Homes.
Holmes Homes has already helped numerous customers access this tax credit and continues to have strong interest from potential buyers looking to capitalize on this buyers' market.
Spencer Holmes, Partner in Holmes Homes states, "It has been incredible to see the response of our customers as we have helped them to understand and acquire this tax credit. With only a few months remaining, we continue to see potential home buyers coming in and wanting to be a part of our program, the "Federal Tax Credit Advantage."
Potential home buyers can visit any of Holmes Homes 9 decorated models in Daybreak for more information and program specifics.
$8,000 Home Buyer Tax Credit at a Glance
- The tax credit is for first-time home buyers only.
- The tax credit does not have to be repaid.
- The tax credit is equal to 10 percent of the home's purchase price up to a maximum of $8,000.
- The credit is only available for homes purchased on or after January 1, 2009 and before December 1, 2009.
- Single taxpayers with incomes up to $75,000 and married couples with incomes up to $150,000 qualify for the full tax credit.
- Homebuyers can "amend" their 2008 tax return to receive an expedited cash benefit of this tax credit.
For additional information, please reference the National Association of Home Builders' website for a complete list of details and answers to some of the question you may have about this opportunity (http://www.federalhousingtaxcredit.com)
Holmes Homes is a 5th generation builder that is committed to building only the most energy efficient homes in premier locations. More than 16,000 homes have been constructed by the Holmes Family who builds Utah townhomes, condominiums, and single family homes. Aside from home building, Holmes Homes and its subsidiaries are involved in land development, home financing, and real estate brokerage operations. Thus far, Holmes Homes has constructed nearly 700 Daybreak homes alone and will soon be celebrating its 120th anniversary in the coming months.


text:
BLOW MOLDING MACHINE Heavy Duty Blow Molding Machine Contact Horng Ying Application Engineering Department For The Wide Range of Moulds
This machine is specially designed for: Washer large and small type of seal iron shell Features: Drug liquid adhesive uniformity dryer very quickly and completely. The operation adopted automatic/hand control model for easy operate. Adopt the air-drive diahragm pump for drug liq
BLOW MOLDING MACHINE HY650 Single Die Head, Single Station Accepts PE, PP, PS, PC,PET-G, Nylon Materials Computerized Control
Featured Products
BLOW MOLDING MACHINE/Fully Electric Blow Molding Machine/Plastic Processing Machine/Injection Molding Machine (HY-900E~1200E)
PVC tile back cementing machine


text:
Other talks from Dhammakranti
There are no tracks or transcript for this talk. Help us make them!
Falling on the full moon day Subhuti's talk observes the Indian tradition of giving talks on this day, as requested by the wife of the patron of the Siddhart Vihara, Gulbarga where the retreat was led. We hear the story of Ajatasatru which then leads into the importance of meditation and its place in the spiritual life.
|Dhammakranti Gulburgha 4: Full moon day and Meditation (57:03)
Total running time: 57:03


text:
September 06, 2016 10:00 ET
ST. JACOBS, ONTARIO--(Marketwired - Sept. 6, 2016) -
Editors Note: There is a photo associated with this Press Release.
The much anticipated Canada 150 tulip has left Home Hardware's Distribution Centre in St. Jacobs, Ontario and is making its way into all Home Hardware Stores across Canada. Officers with the Waterloo Regional Police Service were on hand to help kick off the tulip's trek. This limited edition bulb, created as the official tulip of Canada's 150th anniversary is available exclusively at Home Hardware in limited quantities.
Home Hardware Stores Limited has partnered with the National Capital Commission (NCC), the official gardener of Canada's Capital Region, the Kingdom of the Netherlands and Communities in Bloom for the selection and distribution of the Canada 150 tulip.
Developed in the Netherlands to celebrate Canada's milestone in 2017, the tulip is white with red flames and bears a striking resemblance to the Canadian Flag.
"As a 100% Canadian owned and operated company, we are proud to bring such a unique offering to our customers to celebrate an important milestone in Canada's history. We look forward to seeing the tulip blooming in communities across Canada in 2017."
-Terry Davis, CEO, Home Hardware Stores Limited
About Home Hardware Stores Limited
Home Hardware Stores Limited is Canada's largest Dealer-owned hardware, lumber, building materials, and furniture cooperative with close to 1,100 stores under the Home Hardware, Home Building Centre, Home Hardware Building Centre and Home Furniture banners with annual retail sales of over $5.8 billion. Founded in 1964 in rural St. Jacobs, Ontario, Home Hardware remains 100% Canadian owned and operated. Through the Home Hardware network, Dealer-Owners have access to 100,000 quality, brand name and private label products, a state-of-the-art distribution system and extensive marketing and training programs. Offering a mix of tradition and innovation, Home Hardware Stores Limited has received designation as one of Canada's Best Corporate Cultures and Best Managed Companies and is committed to providing local communities with superior service and expert advice. More information about the company is available at homehardware.ca.
To view the photo associated with this press release, please visit the following link: http://www.marketwire.com/library/20160906-1068204e_800.jpg
Jessica KuepferPublic Relations ManagerHome Hardware Stores Limited(519) firstname.lastname@example.com OwensPublic Relations CoordinatorHome Hardware Stores Limited(519) 664-2252 ext. firstname.lastname@example.com
See all RSS Newsfeeds
Home Hardware Stores Limited and Waterloo Regional Police send the limited edition, Canada 150 tulips off in style outside of Home Hardware Headquarters in St. Jacobs, ON, Friday, September 2, 2016. The Canadian Press Images PHOTO/ Home Hardware Stores Limited


text:
How to quickly build up a website for yourself
Posted by cfhi at July 15th, 2015
A website is more than a simple page online. It happens to be your online ID. It is also your base for operations when you do transactions, or posting important details about your business and even product promotions. How it appears to the people will really matter, and more than that, the content and how you present it to the public will determine if your site is worth the time and effort to go into.
In the past, you would rather hire web page developers to help you get your own website started. It is a good idea because of course, these people know what they are doing. They will be able to create such a good website for you that you don't really need anything else, just simply post it online and monitor it from that point. The only problem with this setup is that it is very expensive to actually hire a web developer! If you are not aware, it can take thousands of dollars to hire them, from conceptualization, up to the end product which is the website itself, it is not that easy to develop the site with them if you are on a very limited budget. So what is the possible solution for this one? Creating your own website!
How to quickly build your own website? Here are the possible steps to take:
-Research the best kinds of programs or software you can use to develop your website. You will hear a lot of good things like WordPress, Joomlar, and even the Microsoft Publisher, dreamweaver, and so many more. The only problem is that they are not that simple to learn. You have to really dedicate time in order to learn to use it. In the meantime while you are learning how to use or utilize the program….
-Conceptualize the idea of the site you want. Be specific and up to point. What is the website for? Is it for a simple personal web page, or it will be used for business? If so, then make sure that your site will be specific and contain only necessary information that is related to your product, business, or personal.
-Collate necessary pictures, charts, videos, and any other things that you will be needing for your page. It does not matter if you collate so many things and go more than necessary. Remember, these will still be streamlined and selected based on the needs at the very end of the site's development. The more to choose from, the better because you never know which combination will work out well for your page.
-When you choose the right program, make sure that you understand the nuances and advantages of using the program. You see some page creators like wordpress or such might be compatible with windows or not, so predict in advance also what will work well with the platform you are using.
-Web hosting! This is important because after you have created your webpage, it is time to be uploading it on the internet. It is only possible to do that by selecting the right web hosting page. You can select from famous ones like Linode or DigitalOcean, and many more. Be sure to review the rates, know their uptime, and even know how it is they manage your site independently so that you will have all the technical support you need when the time comes too.
-Get help from professionals as needed. You don't have to really hire a professional to do things for you, but if you have friends who know what they are doing, then why not ask them right? A few small tips and tricks can go a long way for your website, and if it is something from professionals themselves, then chances for success and improvement for your page will significantly be higher.
Preparation is always the key for building your own website. Make sure that if you decide to quickly build your own website, you are ready to dedicate a whole lot of time and effort because it will be all for naught if you do this halfheartedly. Creating a website might sound easy, but if you will simply put in anything you like without considering the nuances of a good website, then you are wasting time, effort and money.
If you also need help on this one, then you can even go online and read up on tips and tricks on how to create your own websites. Tutorials are even available in youtube if you want, and the best part is that they are all free. If you are to assess things, you will be surprised to know that a good percentage of websites these days have been created personally without requiring the help of professionals. If they are able to do it themselves, what stops you from doing the same?
Category: Internet Security & Online Privacy


text:
eighty one Topic Ideas For Starting A Weblog That Matters
A weblog and podcast for unbiased creatives and entrepreneurs constructing matterful things. Unhealthy Health IT ("BHIT") is outlined as IT that is sick-suited to function, onerous to make use of, unreliable, loses knowledge or gives incorrect data, is tough and/or prohibitively expensive to customize to the wants of various medical specialists and subspecialists, causes cognitive overload, slows quite than facilitates customers, lacks acceptable alerts, creates the need for hypervigilance (i.e., in the direction of avoiding IT-associated mishaps) that will increase stress, is lacking in safety, compromises affected person privateness or evidentiary fitness, or in any other case demonstrates suboptimal design and/or implementation.
Bear in mind right now isn't all about the delicious meals, however concerning the family you're spending time with and the memories you are gathering in your coronary heart. I dedicate 2 hours a day, 4 days every week, to fitness training and have been doing this for the previous sixteen years. 26. Yang Y, Breen L, Burd NA, Hector AJ, Churchward-Venne TA, Josse AR, Tarnopolsky MA, Phillips SM. Resistance train enhances myofibrillar protein synthesis with graded intakes of whey protein in older males.
Now I would like to return and attempt to discover the place I used the unsuitable phrase. Hello, is it necessary to have a fitness instructor certificates before beginning to write a health weblog? In case your weblog is/turns into a business, you could serve your readers by giving them the most related and useful information. It sort of all clicked together and realized I wanted to take the love I had for blogs and motion it with one thing I'm keen about.
The backrest on the Bowflex SelectTech 5.1 and three.1 is easy to modify to totally different angles to enable you to perform completely different exercises including free-weights, sit-ups, bench press, decline bench press and extra. Thanks for these good health record.. I really like fitness something related to health is so important. A LinkedIn search revealed that more than 20 former Twitter staff at the moment are at health, health and bio-tech start-ups, including former CEO Dick Costolo, media head Katie Jacobs Stanton, and engineering VP Nandini Ramani.
Okay, first up is the widespread strikes that are used to create that beloved tap dancing sound you get when the whole lot is in sync and are actually looking like a Professional whereas using it. Sure, I have been there and have impressed just a few buddies that witnessed me banging this little speed bag thingy into submission, all while faucet, tap, tapping my manner right into a fictitious appearance of a voracious fighter on the free.… Read more


text:
I have knee replacements for both knees now, My question is that the one I had replaced in 2010 is doing well and the recovery time was great but the most recent was in 8-06-2013. on 8-16-2013 they went back into my knee because there were two blood vessels bleeding which in turn filled up my knee cavity and down into the ankle. There has been extreme pain since and is still sore to walk on after 30 minutes or so of being on my feet, my orthopedic surgeon says everything looks good but I don't understand why this surgery is taking so much longer.
Knee replacement - why is one knee taking so much longer than the other to heal?
- Asked
- 30 Oct 2013 by ebccotton
- Updated
- 8 October 2014
- Topics
- knee joint replacement, knee, recovery
Responses (1)
yes it was the same Doctor plus not only does it hurt in the knee area but is also sore in the calve area and not able to sleep during the night no more than 2 to 3 hours unless I take a sleeping aid, and hurts 24/7.
I am assuming the system used was the same. If there was more vascular disturbance it will of course slow the healing process and create more pain. I agree with kaismama's comments. Elevation (well above the heart) should help some what.
Hi Ebccotton,
Did your doctor find out the possible cause of your bleeding blood vessels? Can you rate the pain from 1-10, 10 being the highest? I hope everything is well with your knees already.
Search for questions
Still looking for answers? Try searching for what you seek or ask your own question.
Similar Questions
Knee Joint Replacement - When can I resume taking naproxen?
Posted 1 Jun 2014 • 4 answers
Knee Joint Replacement - My knee replacement destroyed the right side of my body. My thigh, hip, and
Posted 18 Mar 2016 • 3 answers
Knee Joint Replacement - I had bilateral knee replacement 1 yr and 2 months ago, I don't have pain?
Posted 3 Apr 2017 • 1 answer
Knee Joint Replacement - Hi. I had a partial knee replacement 12 yrs ago. The replacement slipped?
Posted 29 Dec 2017 • 1 answer
Knee Joint Replacement - How long till the stiffness subsides in my operative knee?
Posted 18 days ago • 1 answer


text:
Community Spotlight: Bright Water in Snellville GA
Bright Water in Snellville, Gwinnett County, Georgia is a Swim-Tennis Lake community with homes available for sale now! This neighborhood has a great location as it has easy access to shopping facilities, great schools, and medical centers.
Inside Bright Water are great facilities that residents can use for fun or if they would like to join their active teams. There is a 7-lane, 25-yard, swimming pool that residents can use from May to September. During these hours, a life guard is available to watch over the swimmers. The community team is The Bright Water Breeze swim team and they compete in the Gwinnett County Swim League. In 2003, 2008, and 2009, they have won the Division Championship.


text:
All of us learn a great deal through our involvement in Cork Community Bikes, and we are keen to teach others, whether at DIY workshop sessions or in training courses.
We deliver bike maintenance courses throughout the year, in the workshop and in other locations around Cork. Most courses we run are suitable for beginners, but we also teach at more advanced levels.
Our next course will be a beginners' course in bike maintenance. Starting on Thursday 23rd November, 7pm, it will run every Thursday evening for 5 weeks in our workshop.
6 places are available, please contact us as soon as possible to reserve a place.
All tools and materials provided. Cost is €30, please bring payment on first day. The workshop can be cold during winter, so please bring warm clothes.
If you are interested to hear about training courses we run, please join our training list using the form below.


text:
Dr Colin Restall is an Associate Professor on our Biochemistry Degrees.
Dr Colin Restall is an Associate Professor on our Biochemistry Degrees.
This module begins with an introduction to enzymology, enzyme kinetics and enzyme regulation. Next there is an introduction to bioenergetics and a discussion of the central metabolic pathways by which energy from food is used to fuel the synthesis of the energy currency molecule ATP (with particular attention to glycolysis, the citric acid cycle, ß oxidation of fatty acids, and oxidative phosphorylation). The concept of tissue and organ specificity is introduced, and finally the role of the hormones insulin, glucagon, epinephrine, growth hormone, thyroid hormone and leptin in controlling these pathways and producing effects on the body is considered.
This module is designed to develop the skills required for students of biochemistry and medical biochemistry. Students meet with their tutors and will be given a series of assignments designed to develop skills in key areas such as essay writing, presentations and general numeracy. Joint session cover generic skills such as internet searching for scientific sources, referencing and plagiarism
The module provides a broad study of the interactions between chemical processes and biological systems. It addresses the chemical processes used by organisms to utilize chemicals for energy production, structural functions and as building blocks of macromolecules.
The module begins with a discussion of selected metabolic disorders of carbohydrate metabolism and their pathological consequences with particular emphasis on diabetes mellitus. The molecular mechanisms underlying polycystic ovarian syndrome are then considered. This is followed by a consideration of the biochemical basis of haemostasis and blood clot lysis. Next there is a discussion of inborn errors of metabolism, and the biochemical pathways affected. Finally the module reviews mechanisms of protein transport and their relevance to storage diseases.
This module reviews the basic elements of enzyme kinetics, including their mechanism and inhibition. The control of enzyme function by allosteric mechanisms is considered together with an appreciation of the significance of sigmoid kinetics. An overview of coenzymes and (metal) co-factors in enzyme function is given together with a description of the role played by vitamins in co-enzyme production. The module concludes with an overview of enzymes from extremophiles and their applications.
The module reviews the pathways of fatty acid degradation (beta-oxidation) before describing the biochemistry of the multi-functional proteins that catalyse the formation of fatty acids. The mechanisms of fatty acid elongation and desaturation are covered along with the synthesis of triacylglycerols and glycerophospholipids with consideration being being given to the control of these pathways. The synthesis of cholesterol via the mevalonate pathway is described along with the various metabolic control points and an understanding of dietary lipid transport. The further metabolism of cholesterol to bile salts, steroid hormones and vitamin D is outlined with consideration given to the stereochemistry of these compounds. The module provides a sound basis for understanding the metabolism of amino acids, lipids and steroids. The pathways of amino acid synthesis and degradation in humans are described along with an introduction to the urea cycle.
The module will provide a sound theoretical and practical basis to a comprehensive range of biomolecular techniques currently employed in molecular genetics and biochemistry. Particular emphasis is placed on DNA recombinant technology, next generation sequencing, transcriptomics, proteomics and mass spectrometry.
Bydd y modiwl yn darparu sylfaen ddamcaniaethol ac ymarferol gref ar gyfer ystod gynhwysfawr o dechnegau biofoleciwlaidd a ddefnyddir ar hyn o bryd mewn geneteg foleciwlaidd a biocemeg. Rhoir pwyslais penodol ar dechnoleg ailgyfuno DNA, dilyniannu cenhedlaeth nesaf, transgriptomeg, proteomeg a sbectrometreg màs.
In order to help students understand the biological basis for behavioural neuroscience and neurological disorders, this module seeks to integrate the multidisciplinary sciences - for example, anatomy, physiology and biochemistry - that have combined to build the emerging field of neuroscience. The aim is to gain a mechanistic and holistic knowledge of the nervous system that builds from the molecular, cellular and developmental, to the systems level. In addition to exploring normal function, this module will introduce common disorders of the central and peripheral nervous systems in an integrated way. Students will be guided in exploring the scientific evidence around what is known and unknown and will be introduced to current research findings in the scientific literature.
The module provides an understanding of the dynamic structure and properties of biological membranes and the nature of lipid-lipid and lipid-protein interactions. An appreciation of the molecular architecture of energy-conserving membranes is also provided together with an understanding of the molecular mechanisms involved in energy transduction.
Students will acquire a detailed knowledge of one topical research area of key importance to their particular degree scheme.
2011 - Present


text:
Safety Statement
Propose a Method
Cite a Method
Technical Sub Committees
Archived Methods
Laboratory Proficiency Program
Prefaces to Earlier Editions
Levels of lower boiling volatiles in beer and ale can be determined by several procedures. This method determines the concentrations of ethyl acetate, n-propyl alcohol, i-butyl alcohol, the i-amyl alcohols (composed of i-amyl and active amyl alcohols), and i-amyl acetate in beer or ale by direct injection of a beer sample containing an internal standard onto a gas chromatographic column. The concentration of each of these compounds is obtained by calculating the ratio of the unknown peak to the internal standard and comparing this value to a calibration curve.
ASBCnet Home | Statement of Copyright | Terms of Use and Limits of Liability | Contact Us | ISBN 978-1-881696-21-6


text:
"One for the rockstars: a secluded recording studio, with seven immaculate bedrooms attached"
With only 1 villa, 1 suite, and 3 cabins deposited across the property, you really can't feel cramped here even when it's full. The sound of reggae may filter through your day, but you can't be sure if it's nearby local musicians or other guests in the recording studio. All sleekly tropical accommodations have ocean views from their outdoor verandas - some have Jacuzzis, others feature a hammock to complete the delightful lounging experience. There's not much to do here other than practise your cool skills in the Bushbar - or to be a little bemused by the in-room technology: home theatre, digital surround sound, multiple dimmers, hidden speakers, iTV, 10,000 songs, 170 movies. As one guest said, it's "Robinson Crusoe goes to the Apple store". And whilst the hotel has a private beach at Macks Bay, Portland also offers several unspoilt areas for you to explore…if you can be bothered to break the spell.
- Extremely pampered privacy, incredibly exclusive, faultless service
- With the highest rainfall on the island (short bursts only 5-7am) you really get a fix of green lush foliage
- High quality amenities (500 thread-count sheets, uber-luxurious bathrooms) and state-of-the-art technology in all rooms
- Even when full it feels empty, and if you take on all the rooms you can have the run of the place and your own private hotel
- A pretty, secluded cove and beach that extends your luxury lounging right to the waters edge
- The 3 cottages are circular and on the small side (rectangular furniture always looks a little awkward against curved walls) but it's a case of small bedrooms, big views
- The pool is in Sanwood Villa, and if other guests book it you will be locked out
- The property straddles the coast road, you need to cross it to get to the beach, and there can be intermittent traffic noise, but it's hardly a rush hour, and you don't notice if the music is on
Best time to go
Our top tips
- Boutique Hotel
- 1 Suite, 1 Villa, 3 Cabins
- Restaurant + bar
- All ages welcome
- Open all year
- Outdoor Pool
- Spa Treatments
- WiFi
- Pet Friendly
- Disabled Access
- Beach Nearby
- Off-street Parking
- Restaurants Nearby
- Air Conditioning
- Guest Lounge
- Terrace
- Garden
- Gym
- Pool Table
Each of the rooms are bright white and super-slick: white walls, white cabinets and white leather bed heads, dark hardwood Jamaican floors. Grey tiled bathrooms have a shower only, and some double as steam rooms (Mento and Rock Steady) and all look out onto lush green foliage and the sea. Blackout blinds ensure you won't be woken by the dawn. At Geejam luxury is techno heavy: flat screen TV, Apple music, 10,000 songs. There is no room phone, but you are given a mobile on arrival which connects you to the Bar, Chef, and other rooms if required. No in-room coffee and tea making facilities but these are available on request. Whichever room you choose, all have lovely dark timber decks, with cane furniture and plenty of cushions to help you break your stride.
Families are encouraged to stay in Sanwood, the main 2-storey villa, which is the original building on the property - designed by British architect, Anthony Wade, in the 1960s. The villa has 3 bedrooms all with ensuite shower bathrooms, 2 terraces, a snazzy media lounge and home theatre, private pool, and, original Banksy artwork plus Juergen Teller photographs - both are former guests. Do note that the Geejam kitchen is located adjacent on the ground floor, so you will see the chef coming and going.
The 3 timber cottages are tucked away in the grounds. Our room, Rock Steady, is the smallest - definitely no room for extra beds here - but it does have a double steam room. Mento has high ceilings and the benefit of an additional hammock on its wraparound veranda as well as its own Jacuzzi. While Ska's double glass doors open onto not one but 2 decks, 1 of which also has a Jacuzzi with a view of the sea.
The signature 1 bedroom suite Drum & Bass is below the recording studio, and has extensive views from its capacious deck.
Features include:
- Air conditioning
- Bathrobes
- DVD player
- Extra beds
- Ipod dock
- Minibar/fridge
- Phone
- Safe box
- Satellite TV
- Slippers
- Terrace
- Toiletries
- WiFi
Food is served in the extensive open-air Bushbar, with its welcoming supersized sofas, ambient sound system, views towards the ocean, and more tables than are really required. The menu is Asian/Jamaican/Caribbean fusion. A set choice covers breakfast and lunch, but the dinner menu is changed weekly. The kitchen wasn't firing on all cylinders on our visit as they had a tiny turnaround window between mega-stars, but what the chef rustled up for us in Sanwood was delicious. As were the mojitos and rum punches...
Breakfast is either Continental with fresh baked pastries, or Jamaican breakfast of the day - steamed dumplings, ackee and salt fish, plantains or banana pancakes. Lunch is a choice of three salads to start, and seven mains including Geejam spicy lobster, red stripe beer batter fish and chips, followed by three puddings that included cheesecake, and ice cream. Dinner offers four starters and five mains; enjoy the chef salad, followed by escovietch fish, curry shrimp, and the Geejam brownie and ice cream to finish.
Sanwood has its own dining options - you can order from an a la carte menu - the fully equipped kitchen adjacent prepares food for all guests, which is taken over to the restaurant for final cooking and assembly. Bespoke menus and room service are available on request. Guests staying in Sanwood would not cook there as all meals are prepared by staff. Welcome baskets are not a standard service but can be requested at an extra cost. Guests renting Sanwood with specific food requirements can be accommodated.
As an alternative you should head over to Boston Bay - the birthplace of jerk seasoning - at some stage for the original jerk pork or chicken, and a catch-of-the-day dinner can also be arranged at one of nearby Port Antonio restaurants. Geejam also recommends Italian owned San San Tropez or contemporary Jamaican Norma's at the marina.
Features include:
- Bar
- Minibar/fridge
- Restaurant
- Room service
- Stroll down (for 3 minutes) to the recently created private beach area at Macks Bay, to swim, snorkel (the hotel has gear you can borrow), or take out a special glass bottomed kayak
- Check out the beautiful local beaches: bright white Frenchman's Cove (10 min walk), incredible, San San Beach, Boston Bay, Long Bay (stunning beach but strong swimmers only)
- Head over to the Blue Lagoon, a few mins drive or 20 min walk, and experience where warm ocean water meets cold spring water
- Raft on Rio Grande - a very popular trip, and have a home cooked lunch on the riverside
- Fulfill that music fantasy with the Recording Studio Experience, details on request
- Check out Port Antonio - the nearby historic coastal town, to see the Errol Flynn Marina and heritage buildings; at night check out its reggae sound systems and ska bars
- While there arrange to have some scuba diving tuition
- Chill on the Day Dream Canopy Deck and gaze at the waterfall
- Have a Spa treatment on the veranda above the recording studio or in your room - choose from a wide choice of massages (Swedish, lymphatic drainage, pre-natal, deep tissue, and Geejam's own tropical touch), or have a facial, mani and pedi
Activities on site or nearby include:
- Historical sites
- Kayaking
- Music
- Nightlife
- Plantlife / flora
- Pool table
- Rafting
- Scuba diving
- Scuba diving courses
- Snorkelling
- Swimming
- Well being
Children are welcome and accommodation is free for children under 12 who share a room with an adult. Babysitting / nanny service can be arranged on request.
Best for:
Children (4-12 years), Teens (over 12)
Family friendly accommodation:
All rooms can accommodate baby in a cot. The 3 cottages are too small for extra beds, and families are encouraged to stay in 3-bedroom villa Sanwood, where you get the bonus of the pool. Drum & Bass can take a rollaway bed.
Families Should Know:
Kids would have great fun playing in the grounds, but you need to cross to main road to get to the man-made beach: sandcastles are possible, but if you want to play in the sea you need to go down the road to Frenchman's Cove.
For more family-friendly places, see our Kids Collection


text:
What is GeeqChain?
GeeqChain is the most scalable, secure and flexible blockchain platform on the planet.
GeeqChain uses a highly efficient, dynamically adjusting, hub-and-spoke network which allows it to handle arbitrarily large transaction volumes.
GeeqChain is based upon a new Proof of Honesty ™ validation protocol, which provides Strategically Provable Security™, effectively, 100% Byzantine Fault Tolerance.
GeeqChain is designed to support an ecosystem of federated chains that can exchange tokens and share business processes.
GeeqChain Explained
GeeqChain is a scalable, inexpensive, and computationally light approach to building blockchains called the Catastrophic Dissent Mechanism (CDM). The CDM is based on game theory and economic mechanism design and takes a completely new approach to transaction validation: Proof of Honesty (PoH).
What is Different About GeeqChain?
GeeqChain has had the benefit of learning from Ethereum which, in turn, had the benefit of learning from Bitcoin. As a result, the GeeqChain platform has been able to solve the most significant outstanding problems facing blockchain today.
Designed for Scalability
GeeqChain's hub-and-spoke network is designed for transaction loads that exceed the combined capacities of the Ethereum and Bitcoin blockchains, by orders of magnitude.
The Most Secure Blockchain on the planet
GeeqChains are the only blockchains that offer 100% Byzantine Fault Tolerance. Our validation protocol, Proof of Honesty (PoH)™, paired with our platform, creates a system such that no individual, or group of co-conspirators, has any incentive to be dishonest on blockchains within the GeeqChain ecosystem.
Unprecedented Transaction Flexibility across Blockchains
Two of the most common questions asked by people who are curious about cryptocurrencies and customized tokens are: "What makes one cryptocurrency better than another?" and, "Why should I think about customized tokens?". Those are exactly the right questions to ask and, in fact, the answers are interrelated.
Be a Geeq.
- or -
For more information on how to customize GeeqChains as a service, please contact: email@example.com


text:
Use this file to export/import wireless network info (especially PSK's) to different machines. This seems to be most helpful when using PSK's that are lengthy to avoid typing them on multiple machines.
List the available wireless network profiles:
netsh wlan show profiles
Or, Control Panel > Network and Internet > Manage Wireless Networks
netsh wlan export profile="NAME" folder="DESTINATION" key=clear
NOTE: by specifying the parameter "key=clear" the PSK will be PLAIN TEXT in the XML file, protect it accordingly!
Command prompt:
netsh wlan add profile filename="FILENAME" user=all
Using the command above in a batch file will allow some automation. I copy the batch and XML files to a flash drive to enable easy adding of the network to a newly imaged computer. You can also drop this into login scripts or other automation tools you may have.
Now that the batch file is created you can use other tools to automate the deployment of the wireless settings. Again, keep in mind that the key is stored PLAIN TEXT in the XML file.
Yeah, I mucked up and stumbled on profile="Name". I thought it meant the user profile, not the wifi network name. On a lighter note just entering: "netsh wlan export profile" will export all stored profiles as separate XML files.


text:
Robert D'Andrea, convicted in 1993 murder, heading back to prison
A convicted killer who served time for murdering his friend in a Hicksville park 23 years ago is now heading back to prison.
Robert D'Andrea pleaded guilty Wednesday in Mineola to having a loaded, illegal handgun and heroin in his car when he was pulled over in Oceanside back in May.
He had been released on parole after serving 10 years behind bars for the shotgun murder of Bryan Levine, 16, who was found dead in Libby Park.
Nassau Homicide Detective Michael Kuhn picked up the cold case and arrested D'Andrea in 1999 for the 1993 murder. He says it took six years because, at first, none of the neighborhood teens would talk.
"As they got older and more mature, they realized how heinous of a crime this was," says Kuhn.
D'Andrea, who was 15 years old at the time of the murder, had been sentenced to seven years to life.
On the new charges, a judge promised him five years behind bars when he is sentenced in September.
D'Andrea's legal aid attorney declined comment about the case.


text:
- Our advice is free and impartial, even if you go elsewhere after speaking to us
Insurance made simpler with LARK Private Clients
A potential client called our Farnborough office, asking us to quote on his home insurance.
He was worried that he'd be wasting our time. It was, he feared, all too complicated. He was struggling to get a reasonable quote online as his answers didn't fit the insurers' "boxes" and the companies he called just couldn't help.
Our dedicated team understands complicated homes; they excel in those properties that are different, for whatever reason. Old properties, unusual properties, properties deemed "non-standard construction" including thatched, timber frame, stone, flat roof, windmills, watermills, converted barns and high value homes.
As long as it is your main home of residence or your main home is insured with Lark, we can help!
Even on the rare occasion that cover cannot be placed, you will still receive our best advice.
In addition, our advice is free and impartial, even if you go elsewhere after speaking to us. So at the inception, adjustment or renewal of your mid or high-value home, you will not be charged.
You won't be wasting our time, it's our job and we're passionate about insurance and doing the right thing for our clients. Don't wait for renewal; speak to a member of our team now


text:
Stream your favorite radio stations with TuneIn and listen to the latest NFL, MLB, NBA, NHL and NCAA college football and basketball games live. With 100,000+ AM and FM stations, TuneIn offers a wide selection of music, news, podcasts, audiobooks & sports radio from around the world.
Download TuneIn Radio and listen to all of your favorite programming for free and on-demand!
Top 5 Reasons to Download TuneIn:
- Stream a wide selection of AM & FM radio stations with varied programming from around the world
- Listen to live sports coverage, including NFL football, MLB playoffs, NHL and NBA games
- Listen to your favorite talk shows and podcasts live and on demand
- Download your favorite audiobooks and podcasts for offline listening
- Hear every play in your favorite NCAA college football and basketball games, including live CFB and March Madness broadcasts on Westwood One
With the free TuneIn Radio app, you can listen to your favorite radio stations anytime, anywhere. Stream motivational music from your Apple Watch while you workout, or tap into TuneIn for CarPlay and listen to the latest sporting event, podcast or audiobook on the go. Access TuneIn Radio at home with Google Chromecast and enjoy the largest selection of programming from around the world!
Subscribe to TuneIn Premium for even more great content:
- Get live play-by-play from every MLB, NFL, NHL and NBA game, all season long
- Let your imagination run wild with access to a library of 80,000+ audiobooks
- Enjoy exclusive access to 600+ commercial-free music stations
...all with no banner ads!
Subscribe to TuneIn Premium through the app. If you choose to subscribe, you will be charged a monthly subscription fee according to your country. The subscription fee will be shown in the app before you complete the payment. Your subscription will automatically renew every month at the then-current subscription fee unless auto-renew is turned off at least 24 hours before end of the then-current subscription period. Your iTunes account will automatically be charged within 24 hours prior to the end of the then-current period. The subscription fee will be charged monthly. You can turn off auto-renew at any time from your iTunes account settings.
Download TuneIn Radio and listen to varied programming and a wide selection of music, sports, news, podcasts, audiobooks and talk radio shows from all over the world!
Privacy policy: http://tunein.com/policies/privacy/
Terms of use: http://tunein.com/policies/


text:
Audit & Accounting
Financial Planning
Practice Management
Audit & Accounting
Financial Planning
Practice Management
search goes here
Michael Imhoff
Michael Imhoff is a Managing Director of Fixed Income Capital Markets for Stifel Nicolaus & Co. His responsibilities include Municipal Institutional Sales, Trading and Syndication, managing new issue pricing and secondary trading and distribution of tax-exempt debt securities. Mike serves on the firms Board of Directors as well as the Commitment and Credit Committees. Prior to the firms merger with Legg Mason Capital Markets in December of 2005, he was also responsible for all Fixed Income Capital Market management. Stifel, Nicolaus & Co. is a full service Investment Banking firm with 297 offices in 41 states.Mike served on the Municipal Securities Rulemaking Board from March 2006 thru September of 2009. The MSRB was established in 1975 by Congress to develop rules regulating securities firms and banks involved in underwriting, trading, and selling municipal securities. He Chaired the MSRB Regulatory Review Committee and the Finance Committee. Mike is a past Director of The Bond Market Association (now SIFMA) and a past Chairperson for The Regional Advisory Committee from 1999-2000. He serves on the Associations Municipal Executive Committee and formerly on the Regional Advisory Committee from 1999 -2006. He has been a member of the Colorado Municipal Bond Dealers Association since 1985. Mike was appointed by Governor Bill Owens on June 17, 2004 to serve on the State of Colorado Venture Capital Authority and previously served as the Chairperson for the Authority. He serves on the University of Colorado Foundation Board, the University of Colorado at Denver Business School Advisory Board, Boy Scouts of America DAC Board and Colorado Concern. Mike also recently served on the Colorado Infrastructure Finance Taskforce and the Board of the Metro Denver Sports Commission. He is a past Co-Chair of the Childrens Hospital Annual Gala Corporate Committee and American Ireland Fund Gala.
Like what you see? Make sure you're getting it all
Accounting Today is a leading provider of online business news for the accounting community, offering breaking news, in-depth features, and a host of resources and services.
Accounting Today
About Us
Contact Us
The Magazine
Site Map
RSS Feed
Privacy Policy
Subscription Agreement
Content Licensing/Reprints
Advertising/Marketing Services
© 2018 SourceMedia. All rights reserved.


text:
IOI Corporation Berhad (IOI) wishes to announce that we have decided to withdraw the challenge proceeding against the RSPO Board's suspension decision in the Zurich District Court of Switzerland. The withdrawal of the challenge proceeding will be formalised during a conciliatory hearing before the Justice of Peace on 14 June 2016.
Since the filing of the challenge proceeding, IOI has engaged with many of our stakeholders such as customers, NGOs and RSPO to resolve this matter. As of to-date, IOI has completed the Action Plan and submitted to RSPO on 30 May 2016 a Memorandum on our sustainability policy initiatives and resource deployment which was endorsed by our Board of Directors. One of the key policy initiatives stated in the Memorandum is our commitment to voluntarily adopt the more stringent RSPO Next certification system starting from end of year 2016.
The above-stated initiative affirms our commitment and support towards the sustainability principles and the work of RSPO, an organisation of which we are a founding member and on whose Board we have been serving for 10 years already.
Dato' Lee Yeow ChorGroup Chief Executive OfficerIOI Corporation Berhad


text:
- Stop telling yourself that there's something wrong with the way that you look. There's no reason to be looking in the mirror every day and telling yourself that you're fat, that you're ugly, that your skin isn't clear enough, that your hair isn't straight enough, that you don't have the right curves in the right places, or anything else negative. When you tell yourself those things it just reinforces them, so even if it's hard you need to try to cut out the negative thoughts.
- Stop worrying so much about being seen without makeup. You're beautiful even without your face on, girl. After I started participating in makeup-free Mondays I started feeling a lot better about people seeing me without makeup in public again. People don't really care as much as you think they do about the way that you look, and if you stop obsessing over it then you'll feel a lot better about those times you have to rush out of your apartment without any makeup on and without your hair fixed
- Share your insecurities with someone. Even if you feel like they're going to laugh at you, even if they do, it will help a lot just to get those thoughts out of your body and into the air. What's they're out and someone tells you that you're beautiful anyways or that they don't see that problem it's easier to feel better about yourself. I shared some insecurities of mine in my makeup-free Monday post.
- Don't compare yourself to anyone else. That includes your friends, your sisters, random girls you're friends with on Facebook, and even celebrities. You're not them and you're never going to be, so you should start focusing on yourself. They aren't going to be you either, after all.
- Start eating right and exercising regularly. I know this probably sounds ridiculous coming from me, because I'm 20 weeks pregnant and getting big and I can't really do that much to work out, but I do indeed do yoga and I've been eating better than I ever have before even though it's really hard. Just don't starve yourself, make healthy choices, and come up with a workout plan that works for you. It'll make you a lot happier to be healthier.
- Be present in the moment. That means don't be dwelling on things that you've done or things that happened to you in the past as well as don't be constantly stressing yourself out about what is going to happen in the future. It's important to be happy with who we are right now more than who we used to be and who we are becoming.
- Follow a beauty routine in the morning and evening. It's definitely an accomplishing feeling when you get done with everything you need to do before you go to bed or after you wake up. Though I've changed it a little since I made the post at the beginning of the month, I follow a very strict nightly routine before I go to sleep and I wake up every morning feeling happier about myself than I did before I started doing it when i fell asleep every night with makeup on watching TV with my sister or boyfriend.
- Cut negative people out of your life. If you don't live up to someone's standards for how you should be then you don't need them. If they're constantly making rude comments, laughing at you, or adding any negativity into your life then tell them that they need to change it. If they still don't then you probably shouldn't be spending time with them right now. It should be important to your loved ones that you're happy, and happy with yourself.
- Don't dress for other people. I know we've all been through that phase where we strictly follow trends or we dress how all the girls at school dress, but that may not be how we're most comfortable looking. It's important for self-esteem to develop your own personal style and disregard the standards of others. If people don't like you for who you are no matter how you dress then you should just number eight them.
- Do the things that make you happy. We shouldn't be brought down by doing things we don't love doing. Just stick to what makes you happy, it'll make you a lot happier!
Saturday, July 20, 2013
10 Ways to Feel Better About Yourself
9 comments:
Hey girl. So agree with #8. I actually think that brings you down in so many ways, and it's "catching". I also think feeling secure/happy with yourself comes with age. I am sooo ok with myself now than in my teens/twenties. Hell, I started a blog! Haha! Keep up the positive messages that you promote.ReplyDeleteReplies
Thanks, dear (:Delete
Love this positive post, and definitely agree with your tips :)ReplyDelete
Love this post! Good advise!ReplyDelete
Love this post. Good insight and info!ReplyDelete
Such a great post. My favorite way to feel better about myself is looking in the mirror and saying "You look awesome and you will be awesome today." Kind of cheesy but it makes me feel better about myself.ReplyDelete
Basically everything to do to make yourself feel better is cheesy, haha.Delete


text:
Contact by email at:
or by phone at: 641-919-8910
"I'm so glad you found our site and I would be delighted to connect
About Jeannette
Jeannette Koczela spent several decades as an Impressionist oil painter, exhibiting her colorful paintings all over the country and working as a free-lance artist. A growing interest in computers led her to become a graphic and web designer. These interests combined her talents and skills for many years.She created several online products including Wattles Wisdom ArtCard Set, coaching programs, a home study course, and published the book, "Money Mindset Makeover."
Then she discovered her passion for life coaching and after a decade of casually coaching colleagues, she decided to train to be a certified spiritual life coach. As a life coach, she has taught people to partner with Spirit and raise their vibration to a place of empowerment so they can attract more ideal clients and grow their business.
As she began to look for ways to market her coaching business, she looked for a directory to be listed in and a coaching organization to belong to. When she couldn't find what she wanted, she decided to create an association that combined an online directory with a professional industry-standard organization, and thus the International Association for Professional Life Coaches™ was born. She now works full-time on the association to help more life coaches become successful and make their businesses more profitable through more visibility, credibility, and connection.
Her second book, "Life Coach Business Blueprint," teaches practical strategies and tools for life coaches to run their businesses with more profits, success, and enjoyment.
After realizing that almost all of her business was coming from LinkedIn, she developed the program, "LinkedIncome Blueprint," which teaches coaches who want to get more clients how to use LinkedIn to build their own online community that they can market to exclusively. Her third book, "The 7 Essential Steps To Get More Clients" answers the most common questions coaches have about marketing.
Visit her personal website at www.JeannetteKoczela.com
About the International Association of Professional Life Coaches®
Our mission is to inspire, promote, and support life coaches around the world so they can coach the people they were meant to serve and run a profitable business, and so that we can collectively heal the planet and transition into an era of global peace and prosperity.
The International Association of Professional Life Coaches® (IAPLC) is an organization for life coaches who want to get more visibility, credibility, and extra support to make their businesses more profitable. An effective way to establish credibility is to belong to a professional organization. Every industry has them. You've probably seen ones like National Association for Professional Accountants or Association of Professional Architects, etc. When a life coach joins this association, they will be able to show that they are a professional, have more credibility, gain more exposure and visibility in the marketplace, and Gold Members have an additional place to showcase their business in our online coaches directory.
We offer monthly coaching calls that cover various business skills, techniques, and guidance taught by expert life coaches. We also have an online directory with members listed in over 20 different categories. Members listed in our online directory must meet certain criteria to be listed, including credentials, experience, and testimonials.
While there are many organizations for specific coaching training and certifications, the IAPLC is for life coaches with various certifications. Gold Members are given the opportunity to participate in telesummits, giveaways, group publications, webinars, and teach master classes. Bronze Members receive business tools, live-only access to masterclasses, and the IAPLC newsletter.
See what our members are saying HERE.
Call 641-919-8910 for more information on our life coaches from the IAPLC life coach directory.


text:
Contact An Attorney
Your case matters. We will make sure that you are taking the proper steps to protect your rights and your case. The longer you wait, the more your case could lose value. Don't wait! Take action! Contact our office today at 1-888-355-5552 or Contact Us online for a FREE CONSULTATION. We are here to help.


text:
I'll look into it..
But right now i don't have much time but as soon as i got some free time i'll add this feature for sure...
And i would also like to request to other users to give suggestions for useful and dynamic features in this thread. i'll try to add that ASAP.
|lollipop, marshmallow, material design, xposed
|Thread Tools
|Search this Thread
|Display Modes
Linear Mode


text:
How Securus Technologies Closed a Criminal Investigation
Trying to close a cold case can take me and my team days, weeks, or even years, all dependent on the leads and cooperation we get during that investigation. I was in charge of working one case where the suspect basically vanished into thin air after jumping bail, and his new identity meant he could be here in the city or thousands of miles away. I was getting heat to get this case closed, so I turned to some unconventional methods to get help.
The team at Securus Technologies had recently installed a new inmate call system in our local prison, and it was about time I was trained to use the LBS software to help in the investigation. The CEO of Securus Technologies, Ricard Smith, says that his 1,000 employees out of the Texas-based company all have but one objective, making this world safer for us all. We were going to see if that was true.
Once I was able to understand how the inmate call system could help, it made things easier. The software that monitors chatter between inmates on the phone does the work of dozen of corrections officers, identifying key verbiage that is needed in certain investigations. No sooner did we input the name of our suspect, we started to get hits. The covert alert feature pointed us in a very unlikely direction. It appears our suspect had his name changed but he was working for a landscaping company here in town.
The Securus Technologies system detected talk between a relative who was inquiring with his family about the suspect, not thinking anyone was listening or really caring about him. That tip lead us to the company early one day where we found our suspect working on a property, needless to say he was escorted to prison where he sits today.


text:
When Ebola ravaged West Africa in 2014 and a devastating earthquake hit Nepal in 2015, U.S.- based company, NormaLyte, saw an opportunity to make a difference. NormaLyte was created with the goal of producing a great-tasting solution to dehydration. But as the creators continued with its development, they discovered that the ease of portability of their powder product provided an opportunity to do good in parts of the world where dehydration kills countless individuals with no access to IVs. This vision, coupled with Project HOPE's proven track record of effectively responding to disaster, has led to an unstoppable partnership between HOPE and NormaLyte.
Recently, NormaLyte decided to expand their support of Project HOPE by forming a team for the Kentucky Derby Festival Marathon/miniMarathon in April. The team will participate as part of Project HOPE's iHOPE Team, a community of athletes whose passion for a challenge is matched by a passion for Project HOPE's mission. NormaLyte founder, Sam Lee, has personally committed to recruiting 30 runners with a goal of raising $15,000 for the Project HOPE mission. And for their part, the HOPE staff is behind them 100% with all of the tools and resources they need to successfully reach their fitness and fundraising goals.
On behalf of the Project HOPE family of staff, volunteers, donors and people we serve, thank you for your support of our lifesaving mission! We wish the entire iHOPE team the best of luck as they train for the event. One step at a time they are making HOPE happen for vulnerable girls and boys, men and women around the globe.
Join the iHOPE Kentucky Derby Festival Marathon/miniMarathon.
Become an iHOPE Hero! Run, walk, cycle or swim wherever you are.
Stay Informed
Get news from the field and updates on how your donations are being put to work.
Join Us
Read and share stories about Project HOPE with your personal network.


text:
Making wine in Virginia is often a thankless pursuit, determined as much by luck as skill in many years. This stretch of rainy days we are in is a prime example; anything not harvested before the rains started, whether it was ready or not, is now deer and bird food. Fortunately much of the season up to this point has encouraged a fairly early ripening, but any stragglers will be waterlogged and/or rotten messes. Lovingston saw exceptionally good luck a few weeks back when the powerful storm that damaged the Lockn' festival and canceled their first day bypassed them by less than a mile and left them completely unscathed. All wine-growing regions have their risks, but Virginia's can be particularly unpredictable and cruel.
It is with this in mind that we want to honor a wine where all the good breaks happened and a wine of uniquely high quality was produced. Plenty of skill involved here as well, of course, but the seasons were much kinder to the winemakers to allow the raw material to do what they wanted. Lush dark color and deep currant aromas in the glass immediately show the intensity of the vintage, and some swirls in the glass gradually pulls out some spice and red fruits. On the palate the texture is silky and polished as a Merlot should be, but does not lack for character, showing lots of energy behind the dark fruit flavors and fine tannins. This is one of those Merlots that has layers of complexity, an indication of 'seriousness' to it that is many times lost in Virginia wines, and one that could even do well for at least another 3-5 years in the cellar.
INSIDER'S PICK: 2012 LOVINGSTON WINERY MERLOT 'JOSIE'S KNOLL' $19.99(WINE ENTHUSIAST 91pts)
Leave a Reply.
The Best of the Best.
We offering free tastings on these wines in the store every Thursday and Friday, and a 10% discount off the retail price through the duration of the day. Come on by and give them a try!
July 2018


text:
Telstra invests $US18m in apps maker Kony
Applications and Ventures Group is Telstra's venture capital division through which the carrier has taken small takes in start-up technology companies such as IPscape and Ooyala. Photo: Rob Homer
Telstra's venture capital arm has made its first investment of the year after tipping $US18.3 million into US-based mobile applications developer Kony Solutions.
The telco has taken an undisclosed equity stake in the business, which offers large corporate customers pre-built, enterprise applications that can be deployed across multiple operating systems and devices.
Kony already counts Hyatt, Konica Minolta and Schweppes among its customers but Telstra will also market and distribute Kony's products to its local client base.
"For Telstra this is a win-win - we are investing in an innovative company and adding immediate value by being able to offer Kony's products to our business and enterprise customers," Telstra Ventures managing director Mark Sherman said in a statement.
Telstra is believed to have set aside about $40 million each year for investments into start-up software and technology companies in a move designed to unearth new capabilities but also protect the telco against the enchrochment of rivals in adjacent industrues.
Last year, the telco took stakes in a string of companies including Silicon Valley based video streaming service, Ooyala, as well as local based start ups such as restaurant booking service Dimmi and cloud computing business IPscape.
Mr Sherman said Telstra Ventures aims to build an investment portfolio of 20-30 companies over the next three to five years.
"Our investments will be among the best emerging companies in the world each at varying stages of development," he said. "Our team will continue to look for investment opportunities which fill gaps or accentuate opportunities for Telstra,"
Sign up for Computerworld eNewsletters.


text:
I was scrolling through my old blog post and I seriously missed how I literally used to write to you guys about everything and anything. So at the end of each week there will be a blog post that has an entry for ever day o the week explaining how I feel and the juicy details of what went down this week. And if you follow my YouTube channel i will also beposting a video at the end of the week on Sundays that has the same thing. overall i just think this will be so funny to take back on and it will hopefully help you guys get to know me a little bit better
Sunday February 18,2017
Today hands down is one of those days where I realize how I need to stop procrastinating. I have my college meeting for fidm on monday and all day today i have been working on my project, and tomorrow i have to drive to my school because i forgot my recommendation letter in my locker. Basically i am so under prepared for monday and it will look so bad if i don't work my last-minute magic, but I'll keep you guys updated. In the other words lets dig into my personal life something that i never talk about because i hate talking about my feelings. I am currently talking to this guy that I have met ( i am not going to say where) and i can seriously slowly fell us slipping apart and its so sad because we were supposed to hang out next week on friday i think or Saturday, i totally forgot what day we had planned. And other than that I am absolutely loving my new computer case it is so cute and i might do a review on it soon because i know that i love reading reviews on products before i buy them, especially if their from sites like Amazon that are low-key sus. I still dont understand the whole aspect of Amazon.
March 3, 2017
So what happened to updating daily raven? Honestly we all know that wasn't going to happen but let me just try to update you guys won what happen in February and today anyway lol. So the thing with FIDM went pretty well and I have my last meeting this month, and honestly I think I will get in. I'm not trying to be cocky or anything but if the councilor thinks my project is A1, my grades are wonderful, and my family can afford it Im pretty set. Right? Okay yeah I am nervous but in the back of my mind I know it's not going to be a problem. For the last hour or more I think, I have been writing blog post because I am honestly so tired of missing days and watching my blog that I have dreamt of literally turn into nothing slowly. And even though this little effort thing has boosted my blog views majorly I also think consistency will as well. March 3rd is a Friday and I am so mad because I had my whole weekend planned out but of course my mother had to be crazy and say I was lying about what I was doing and made me cancel. So I am sitting here beyond angry just writing to you guys. Have you ever had to cancel your plans last-minute? It's honestly so annoying because gradually my friends stop asking me to hang out with them and think that I am making everything up. It also sucks because then people also think that I just don't like them and dont know how to tell them I dont want to hang out when its not like that. Speaking of not being able to hang out, one of my friends in dance invited me to a Cole concert and I am so excited but I dont want to tell her if I can go just yet because I already know my mother is going to ruin it for me, but its in July so hopefully I can go. Omg, I just remembered its in July! When classes for FIDM start omfg I totally forgot! Hopefully I dont have class so I can go.
Then during practice on thursday my hip hop team coach admitted that she hasn't been feeling the team for a while now, meaning that next year unless anther coach can take over their won't be a hip hop team. To say that I am upset is an under statement because I partially feel like its my fault. When she was telling us she said it's because of personal issues and her mental health, she suffers from anxiety but a situation happen months prior that I feel horrible for.
So one day after practice the ex captain came up to everyone asking if they wanted to perform during the pep rally. And before this everyone was saying how they don't like the choreo, including myself. So of course I said I don't like the chores so no I dont want to preform but I didn't mean it so literal! Do you know how many times I didnt want to do something but I have to do it anyway because its my responsibility and after its done, I'm so happy and proud that the unwell to do the task is far gone. As well as this I also told the ex captain ( because we were friends at the time) that I felt like the coach wasn't taking us seriously because of how many times practice was cancelled last-minute but her other teams practice resumed, and that she always choreographed on the spot. If a coach did that wouldn't you feel the same way? I admit that I shouldn't have been talking about my coach like that because it's just disrespectful but I was just ranting, it's not like I actually meant it to hurt her feelings. Well soon enough I did just that. A couple of days later we had our basketball performance. As always I was so hype cracking jokes until the ex captain came up to everyone with the same drama and asking the same question. Me not taking it seriously said i dont want to preform ( like an idiot) and before I knew it she was going back to the coach to tell her.
Their we are 5 minutes before we go on having a meeting about how coach knows what we said and how for now on she wouldn't be choreographing, as well as if we don't want to perform we dont have to. So everyone including myself who said they didn't want to preform was like, yeah I want to preform, that's what I am on the team for. And their was the ex captain feeling betrayed, but no one told her to tell coach that in the first place, ESPECIALLY FIVE MINUTES BEFORE WE GO ON. So here hip hop team is feeling like shit while the other half is confused as to whats going on. Later the ex captain goes in the bathroom crying or something talking to her best friends and sister, then when I was going to come in I hear he sisster say now you know who your real friends are or something along those lines. And next thing I know I am being called fake for not telling coach that her chororeo sucked, but in my opinion… who was the ex captain to say anything in the first place/not tell her that this was her intention to tell coach…..
So even though we still preformed, while everyone felt like shit because we just all argued and made our coach cry five minutes prior we still did somewhat good. But ever since then the team hasn't been the same, and I still feel horrible for my coach because I should've kept my mouth shut. So now with this information coming out that she feels extra anxious during practice and with this team I can't help but feel like its me who made her feel like crap or that way, and I can honestly never forgive my self for that. And honestly just typing this makes me tear up because I know I seriously hurt her, or contributed to it.
But wow that got deep real fast. So yeah I'm just sitting here blogging about my boring day/reliving terrible memories on a Friday night…. I cannot wait until High School is over!


text:
Description: An architect senses impending doom as his half-remembered recurring dream turns into reality. The guests at the country house encourage him to stay as they take turns telling supernatural tales.
Director: Alberto Cavalcanti, Charles Crichton
Writers: H.G. Wells (original story), E.F. Benson (original story)
Stars: Mervyn Johns, Michael Redgrave and Roland Culver
Year: 1945
Tags: top movie, mystery, Dead of Night
Watch full movie/film online for free and in the best quality here:
*NOTE: If you have a slow Internet connection, choose the lowest film resolution from the menue to have it going faster.
Copyright © 2015 Retro Films .in - Free and structured online films and movies!. All rights reserved.
Last Review 13.01.2015


text:
Canara Bank
Loan Against Property : If you own a valuable asset in the form of a self-occupied house or a commercial property you can go for a loan against property. It is a secured...
Home Loan : Home loan is given for construction / purchase / repairs / additions / renovations of residential house / flat including the purchase of land and...
Can a fixed rate loan be converted to a floating rate loan and vice versa? :Yes, you can convert floating rate home loan into a fixed rate one with no extra charges. However, banks charge a small fee for converting a fixed rate product to a variable rate product. The swap can be done any number of times and at any point of time.
What is a fixed rate loan? :In a fixed rate loan the interest rate on the loan charged by the bank is constant over the tenure of the loan. You should go for a fixed rate only if you feel that the rate of interest prevailing in the market have touched rock bottom and the rates can only move upwards.
How to select the right bank for home loanOwning a house of own is a dream for every person. Lots of people go for any bank loan in the case they can not afford to pay the full price of a house. Banks and other financial institutions offer home loans to people who meet the required eligibility. Various banks offer home loan with their...
The basics of business loanBusiness loans are provided by various banks to business people for their short or long term financial needs. A lot of times it is important for businessmen to acquire a certain amount of money for running their enterprise. It is well known that without the required capital no business can run....
Life Insurance Corporation of India
Life Insurance Corporation of India (LIC) is an old life insurance company in India. It is also the insurer with the largest customer base. The company was formed on 1st September, 1956, with the...
Life Insurance: Children Plan : Jeevan Kishore, Child Career Plan, Jeevan Chhaya, CDA Endowment Vesting At 21, Marriage Endowment Or Educational Annuity Plan, CDA Endowment Vesting At 18, Komal Jeevan, Jeevan Anurag, Child Future Plan, Educational Annuity Plan, Child Fortune Plus
Life Insurance: Retirement Plan : Jeevan Akshay, New Jeevan Suraksha - I, New Jeevan Dhara - I, Jeevan Nidhi
Life Insurance: ULIP Plan : Fortune Plus , Profit Plus , Market Plus I , Money Plus - I
National Insurance Co. Ltd.
Incorporated in 1906, National Insurance Company Limited (NIC), became a subsidiary of the General Insurance Corporation of India (GIC), a Government of India-owned entity after the General...
© 2014 LoanBoss.com | ccentium


text:
Your Download Basket is empty!
Tumbler with Metallic shine on the top.
Branding all around possible..
Fits all 13"-18" laptops
Raises screen level for better air circulation
Better screen level reduc..
- Built- in Single Ultra Bright White LED- Work on 2 x AA batteries - Powerful 3X magnification - 67..
- Set of 4 plastic coasters in round shape, with PU inside
- Classy design, looks elegant with any ..
- Set of 4 plastic coasters in square shape, with PU inside
- Classy design, looks elegant with any..
New improved model, Now works on just 2xAA batteries (not included)3x magnifier with 5 LEDs at the b..
• Made of premium quality Material
• Zipper Insulated Bag to keep Food Warm / Cold
• Preserves tem..
Mr pen holder is a smart way to hold your pens.
Has sticky feet so your pen stays fixed to the ta..
- First of its kind utility design! - Pen with 2 way screwdriver (Both + & -) - Has sma..
• Unique novelty product that everyone would love to keep on their table• Unavoidable, excellent spa..


text:
Ancient NesseburAdd to Cart
Duration: 3 hours
Walking: moderate
Your walking sightseeing tour begins with a visit to the Museum of Art and History, where there are several halls with exhibition items, tracing the rich history of the Old Town of Nessebur. Special interest is risen by the Hall with the icons from Nessebur, which represent the so-called Nessebur's style of icon painting. The facades of the Nessebur's old houses complement the romantic atmosphere of the town and make your visit unforgettable. Walk through the old town, visiting old churches and landmarks. Call at the local restaurant for a lunch consisting of national dishes: "banica", Bulgarian yoghurt, "Shopska salad", different kinds of Bulgarian cheese and other Bulgarian delicacy. After the lunch return to the port.
Write a comment
- Required fields are marked with *.


text:
"Clean and cozy inn"(Review)
Pension Shinano, Madarao Kogen
Pension Shinano: Overview
Nestled in the vast nature of Madarao Kogen, Pension Shinano is located a 24-minute drive from JR Iiyama Train Station and a 1-minute walk from Madarao Kogen Ski Resort. Free Wi-Fi is available throughout the property.
Guests stay in dormitory rooms with bunk beds or standard rooms with single beds. Each is fitted with a TV, alarm clock and a clothes rack. Bathrooms and toilets are shared with other guests.
Book Pension Shinano Hotel now!
At Shinano Pension, guests can use the barbecue facility or relax in the shared lounge with a fireplace. Other facilities offered include laundry facilities, lockers and vending machines.
A simple Japanese and western set menu is served at the dining room.
Madarao Children's Book Museum is a 9-minute walk away, while Madarao Kogen Sanpatikku Ski Area is a 10-minute walk from the pension. Lake Nojiri is located a 20-minute drive from the property.
Rooms: 11
Video for Pension Shinano
Check other Madarao accommodation options
Madarao Accommodation: Quick links to hotels near Pension Shinano, Myoko
Takeda Hotel – Akakura Wakui Hotel – Senke Hotel – Hotel Mumon – Red Warehouse


text:
Jacquard Party Trousers
Get the perfect party look with these metallic jacquard trousers (and matching blazer). We've designed a flattering fit with cleverly placed darts for a sleek shape. And that's not all – the pattern was created in-house and the fabric sourced from a French mill. Bring sparkle to the everyday with a…
- Outer 66% cotton 18% polyester 16% acrylic
- Lining 100% polyester
- Dry clean only
- Fitted through waist, hip and thigh with a straight leg
- Sits below natural waist
- Ankle length
- Front pockets
- Faux back pockets
- Concealed hook and bar fastening at centre front waist with zip fly
Jacquard Party Trousers
£110.00 £44.00


text:
Popular Now
Popular Now
Popular Now
Popular Now
01803 865913
+44 1803 865913
Edited By: William Cronon
561 pages, 35 illus
Essays which argues that many environmental strategies are flawed because they leave people out of the picture.
The best kind of book, one that shocks the reader into entirely fresh ways of thinking.--Michael Pollan
There are currently no reviews for this book. Be the first to review this book!
Your orders support book donation projects
my first port of call for ordering books
Search and browse over 110,000 wildlife and science products
Multi-currency. Secure worldwide shipping
Wildlife, science and conservation since 1985


text:
U.S. Senators John McCain (R-Ariz.) and Carl Levin (D-Mich.) sent a letter to General Martin Dempsey, chairman of the Joint Chiefs of Staff, on Friday asking for his assessment on a number of U.S. related issues, including Syria.
"What is your assessment of the costs, benefits, and risks associated with training and arming vetted elements of the Syrian opposition," Senators McCain and Levin asked in the letter. "In your view, could such action alone be sufficient to adequately build the military capability of the moderate opposition in Syria and create the necessary conditions for the Administration's stated policy objective-Bashar al-Assad's departure and a negotiated solution to the conflict in Syria-to succeed?"
The senators asked Dempsey about a variety of issues related to U.S. policy in Syria. The questions were prompted after a hearing regarding Dempsey's re-nomination was held on Thursday.
"Specifically, does the United States have the capacity, using stand-off weaponry that would not require destruction of Syria's air defenses, to significantly diminish or limit the Assad regime's ability to use air power, ballistic missiles and heavy artillery, particularly against areas of Syria under opposition control?" Senators McCain and Levin said. "Would diminishing or limiting the regime's capacity to use such weapons shift the balance of power in a way that advances the Administration's stated policy objective?"
Dempsey has yet to respond to the questions asked by senators. His answers will be used to evaluate whether Dempsey will be re-nominated.


text:
ERIC Number: ED374047
Record Type: Non-Journal
Publication Date: 1994-Jun
Pages: 25
Abstractor: N/A
Reference Count: N/A
Process Induction: Researching Skills Development across the Social Studies Curriculum. Middle Grades.
Peters, Richard O.
This document discusses the use of process induction and the proactive action model to teach research skills across the social studies curriculum. Process induction is the ability of an individual to transfer schema related knowledge and skills to other situations and settings, to use knowledge and process skills in new and diverse instances to solve problems and resolve conflicts. The proactive action model (PAM) can be used to nurture and enhance students' inquiry and discovery in classrooms and at field based research sites. The true value of knowledge and skills acquisition is its transferability. The transferability quality of social studies content, concepts, subject specific knowledge, and process related skills, is what makes it relevant in the lives of individuals. Students in the middle grades, typically grades 5-8, should be involved in a 4 step induction process: (1) acquisition or the accumulation of concepts, facts, and other information and skills; (2) application or using that which has been acquired to enhance learning and enrich intellectual skills development; (3) reinforcement or the continuous application of innate and acquired skills and knowledge to new situations in diverse settings; and (4) refinement or repeated application of knowledge and skills to new situations resulting in an individual's ability to demonstrate competency or proficiency at some prescribed level. The paper discusses directed learning environments, independent learning environments, and application environments, and how they may be utilized in social studies. Contains 14 references. (Author/DK)
Descriptors: Intermediate Grades, Junior High Schools, Middle Schools, Research Skills, Skill Development, Social Studies
Thought Action Network, 6829 Roseland Dr., Corpus Christi, TX 78414.
Publication Type: Guides - Classroom - Teacher
Education Level: N/A
Audience: Teachers; Practitioners
Language: English
Sponsor: N/A
Authoring Institution: Thought Action Network, Corpus Christi, TX.


text:
While touring Germany, Italy, and Spain, I was highly interested in the symbols each country used to signify basic street information. Here in the states, there has been much debate about if the Accessible Icon is legal since it has not been officially adopted as the symbol of access by the DOT or DOJ. While almost everyone will agree that the symbol represents movement, some argue that a new symbol is not needed or can cause confusion. Since I am now interested in exploring the different symbols that exist, I took pictures of the different symbols I came across while traveling.
Accessible Icon Updates:
- DOJ (Department of Justice) verbal approval of the icon during the National ADA Symposium. Read here (May 2015)
- New York State announcement of a slow phasing in of the symbol. Read here (August 2014)
- New Jersey bill in progress. Read here ( September 2014)


text:
Skip to main content
Sign In
vdsstream VDS
Sign In
Wiki Home
Recent Changes
Pages and Files
Announcements Page
Class Picture
HRI- High School Research Initiative
Fun Plates Sp18
Gel/Nanodrop Sp18
Lab Safety Training
Mini Research Write Ups Sp18
ARI Project Plans
ARI Labs Schedule
Meetings w/ Dr. B
Google Calendar
Hours Chart
FPLC schedule
(for Target Discovery)
The Random Walk
Interesting Snippets
Protocols on GDocs
Virtual Screening
Tips & Hints
NDM-1 Beta-lactamase Project
TACC VINA Work
Mentor Tasks ARI
Mentor Pics
Undergraduate Research Opportunities
Support the VDS Stream with a donation!
Rec Letters
Feed to Announcements
VDS Google Website
DrB old links
Aditi S
No tags
Export (PDF)
The Purification and Characterization of Over-expressed Bacterial Protein BL21(DE3)
In recent years the use of recombinant proteins in scientific research has increased greatly, as has the wealth of techniques and products used for their amplification and purification. [1] The techniques of protein expression, purification, and characterization are especially important to the field of virtual drug screening in that they are heavily involved in the wet lab portion of the research. The expression of the protein usually involves genetically modified plasmids that are inserted into human, bacterial, plant or animal cells in which they are overexpressed. Purification of these proteins often involves enzymes such as lysozymes which can digest cell walls and extract the protein. The characterization part of these labs show the purity of the protein and the overall success of the expression.
Characterizing the purified protein in some detail reduces the risk of wasting resources on protein material of inadequate quality. It also provides a means to ensure that different batches of the same protein have similar properties [2]. This lab aims to use the techniques of protein expression, purification, and characterization to obtain a relatively concentrated and pure elution of pGEM-gbr22, a protein found in coral with a distinctive purple color. If methods are followed properly the protein elution should be purple and its characterization should indicate a heavy, pure concentration of protein.
Materials & Methods:
During expression of BL21(DE3), two samples were iced with plasmid pGEM-gbr22, heat shocked at 42 degrees Celsius for 45 seconds, and returned to ice. 200 ul of SOC was added to the solutions, which were then shaken in an incubator for 30 minutes, and spread on separate LB/amp agar plates with colirollers. The next morning, one bacterial colony was transferred to LB/amp media to shake in the incubator for 8 hours. The starter culture was then moved into an LB/amp flask for further growth. Later, the sample was spun to isolate the pellet, and then resuspended in PBS buffer. For purification, lysozyme and cyanase were added to the bacteria, the soluble fraction was filtered through a 0.22 syringe filter PES with a 5 mL syringe, and purified using a combination of batch and column chromatography. NiNTA resin buffer was used as the 1st rinse through the bio-rad and collected as "Waste". A wash buffer containing 0.2 mL imidazole was used for the 2nd rinse and collected as "Wash." An elution buffer containing 2.5 mL imidazole was used for the 3rd and 4th rinse and collected as "Elution 1" and "Elution 2." The Nanodrop spectrophotometer was used at 280 nm and 574 nm (the maximum absorbance wavelength), along with the extinction coefficient and Beer's law, to find the concentration and yield of the protein in "Elution 1". Samples for each step in the process of purifying were collected and labeled for a total of 6 samples. In characterization, the samples taken throughout expression and purification were heated and centrifuged with loading buffer. Sample 1 was centrifuged for 5 minutes at 5,000 rpm to retain the pellet, which was suspended in 200 ul of water and added to 40 ul of loading buffer. 10 ul of loading buffer was added to Samples 2 to 6. All samples were placed into a heat block at 95°C for 5 minutes and centrifuged for 2 minutes at 5,000 rpm. The samples were then loaded into individual gel cells, and subjected to an electric current in an electrophoresis tank. The gel was then cleaned in nanopure water and stained with imperial protein stain on an orbital shaker. After destaining, the gel was dried on the drying bed, documented, and analyzed.
Figure 1:
Ampicilin positive agar control plate with no BL21(DE3) bacterial growth
in the morning on Day 2
after 24 hour incubation at 37 degrees Celsius.
Figure 2:
Ampicilin positive agar experimental plate with colonies of BL21(DE3)
bacteria transformed with plasmid DNA pGEM-gbr22 exhibited ) in the morning
on Day 2 after 24 hour incubation at 37 degrees Celsius.
Figure 3:
Ampicilin negative agar "fun plate" (coughed on) in the morning on
Day 2 after an incubation period of 24 hours at 37 degrees Celsius.
Figure 4:
A large culture of BL21(DE3) bacterial cells, transformed
with plasmid DNA pGEM-gbr22,
in the evening on Day 3 after
spending 24 hours in the shaking incubator at 37˚C and 250 rpm.
(does this have amp?)
Figure 5:
A wet pellet of 0.32g of BL21(DE3) bacterial cells
transformed with plasmid DNA pGEM-gbr22 in the evening
on Day 3 after centrifuging for 10 minutes at 4°C and 5,000 rpm.
Figure 6:
5 micro-liters of Elution 1 of the protein after purification.
Gbr22 protein after washing with 1x PBS and 250mM imidazole solution.
Figure 7:
5 micro-liters of Elution 2 of the protein after purification.
Gbr22 protein after washing with 1x PBS and 250mM imidazole solution.
Figure 8:
Absorption vs. Wavelength at 280 nm wavelength of elution 1 sample. Blue line represents the absorbance reading.
The Beer's Law formula A=Ebc was used to find the concentration. The absorbance value, the extinction coefficient and the cuvette path length were put into the formula. This equation looks like 0.332 = (38850)(1)(c). The concentration wass found to be 8.546e-6 Mol/L. Multiplying this concentration by the molar weight of the protein (25794.2 g/Mol), the yield was found as 0.2204 mg/mL. Since there is 5 mL, the final yield was 1.102 mg.
Figure 9:
Page Ruler Prestained Protein Ladder Product 26616 Thermoscientific used in this lab for gel electrophoresis.
Figure 10:
Gel after drying on Whitman filter paper, covered with Saran wrap. The lane farthest to the left is the protein ladder. Lanes 2-7 consist of samples 1 through 6. Sample 5, the elution wash, shows about 80% purity. The protein is estimated to be about 25 kDa.
There are two enzymes used in the lab that serve to extract the protein from the bacteria. The lysozyme is used to digest the cell wall of the bacteria, and cyanase is used to digest the DNA and RNA in the mixture. The six histidine residues attach at the C-terminuses (end of the protein) to separate the protein from other things in the solution. Nickel metal binds to the Histine residues not allowing the protein to go through the column until the elution buffer is used. Sample one was cell lysate so protein was present. Sample two was SOL
(what is this acronym?)
and a small amount of protein was present. Sample three was a flow through; no protein (the protein being worked with) should have been present. Sample four was wash buffer and no protein should have been present just the other substances in the solution. Sample five was elution one and should only have the protein and sample six should have whatever protein was left in the column. The buffer is supposed to remove everything else but the protein being worked with. While the elution buffer has imidazole that competes with the histidines to elute the protein off the beads. The size of the protein determined for the gel is 25 kDa which was very similar to the size determined in the protein purification lab. The purity of the protein is 80% because there was only one clear, dark band, and a one very faint one. The protein stayed in the filter until the elution buffer was used. The 6xHis tag and the protein stuck to the nickel. The protein sticks to the Ni-NTA beads and the nickel
(Ni-NTA implies the presence of nickel)
so it doesn't flow through the filter until elution buffer.
Possible sources of error for this lab could include inaccurate measurements of chemicals used, as well as not waiting enough time, or waiting too much time, between steps.
The three labs of protein expression, purification, and characterization lead
to the gel that, when analyzed, showed that there was 80% purity. With this data it could be assumed that the labs were successful. Initially, the target protein was inserted in the bacteria, overexpressed, and then purified using elution 1 and 2. The Nanodrop and Beer's Law were then used to find the concentration. In the protein characterization lab the gel showed that Sample 5 was for the most part one dark purple line, hence the high purity percentage. The molecular weight based on the ladder was found to be 25 kDa, which is similar to the concentration found from the purification lab, showing the labs did work. All the steps learned from this lab are very important later on when more target proteins will need to be harvested, characterized, and used for further testing in independent projects. Using virtual drug screening programs like GOLD combined with PyMol, ligands can be found that bind to the protein to make a drug inhibitor and then tested in the wet lab.
(not necessarily for this lab, it is just a model protein whose functionality we don't necessarily care to inhibit)
[1] Graslund, S.; Nordlund, P., Weigelt, J., et. al, Protein production and purification.
Nat Methods
(2), 135-46
[2] Mattaj, I. W. Protein Expression and Purification Core Facility.
(accessed April 16).
help on how to format text
Turn off "Getting Started"