text: Notary Bulletin A Notary's Guide To Oaths And Affirmations By Kelle Clarke on May 27, 2015 Updated 5-13-19. An oath or affirmation is a solemn promise with legal consequences that can be made before a Notary. If one of your customers wishes to take an oath or affirmation, here are some tips. Difference Between An Oath And An Affirmation While both oaths and affirmations are notarial acts that compel a person to tell the truth, an oath is a solemn, spoken pledge to God or a Supreme Being, while an affirmation is a spoken pledge made on the signer's personal honor with no reference to a higher power. Either is considered acceptable, and the choice is left to the signer. Administering An Oath Or Affirmation When administering an oath or affirmation, follow these steps: 1. The person taking the oath or affirmation must be physically present before you. Oaths and affirmations may not be administered remotely by phone or email. 2. The Notary may ask the person to raise their right hand or make another ceremonial gesture before responding, to emphasize the seriousness of the process. While these ceremonial formalities are seldom required by law, they have value in impressing upon your signer the significance of their actions.The Notary then administers the oath or affirmation by asking if the person swears or affirms the truthfulness of their statement. The wording may vary depending on your state (see below). 3. The person taking the oath or affirmation then answers "Yes" or "I do." The person must speak clearly and audibly so the Notary can hear and understand the response. In California, state law provides the following wording for an oath or affirmation, which requires an affirmative answer to one of the two following questions: For an oath: "Do you solemnly state that the evidence you shall give in this issue (or matter) shall be the truth, the whole truth, and nothing but the truth, so help you God?" For an affirmation: "Do you solemnly state, under penalty of perjury, that the evidence that you shall give in this issue (or matter) shall be the truth, the whole truth, and nothing but the truth?" (CCP 2094[a]). Also, when executing a jurat for a signature on a written document, California Notaries must ask for acceptable proof of identity as prescribed under state law (GC 8202[a]). Texas provides the following oath or affirmation wording for a person taking public office in the state: "I _________ (affiant), do solemnly swear (or affirm) that I will faithfully execute the duties of the office of ____________, of the State of Texas, and will to the best of my ability preserve, protect, and defend the Constitution and laws of the United States and of this State (so help me God)." In states such as Florida where oath or affirmation wording is not specified in statute or other guidelines are not provided, the appropriate wording for an oath for a jurat, verification or affidavit may be something like this: "Do you solemnly swear that the statements in this document are true to the best of your knowledge and belief, so help you God?" Administering An Affirmation In states that do not require statutory wording, common phrasing for an affirmation for a jurat, verification or affidavit might be: "Do you affirm that the statements in this document are true to the best of your knowledge and belief?" Tips For Conducting Oaths And Affirmations Never Take Shortcuts: If the document being notarized requires an oath or affirmation, you must verbally administer it — even if the signer attempts to rush you or avoid it entirely. Ask, Don't Advise: If a signer is unclear about the difference between an oath and affirmation, you may explain the differences but you cannot legally advise the signer which one to perform; it's his choice. Honor Your Signers' Beliefs: Since oaths and affirmations are equally legal and acceptable, the Notary should honor the signer's choice and use the appropriate wording and gestures. Speak Clearly: Make sure the signer responds with a clear "Yes" when you administer the oath or affirmation. Because the signer's answer puts them under penalty of perjury, it's important that the signer's response be clearly understood by the Notary. Nodding, saying "uh-huh" or other ambiguous responses are not appropriate. Take It Seriously: Some signers — and even some Notaries — make light of the situation, but it's clearly not the best time to crack jokes. Expect professionalism at all times by being an exacting role model. Record It In Your Journal: If you administer an oath or affirmation, note it carefully in your journal. Once you recognize the basics and have several declarations under your belt, administering them will be a cinch. You can learn more from your state's Notary regulating agency website or your state's Notary handbook. Or check out the NNA's Notary Primers or U.S. Notary Reference Manual Online, which is a NNA member benefit. The NNA's Notary Essentials eLearning course guides you through all your state's requirements. Kelle Clarke is a regular contributor to the National Notary Association. Related Articles Quiz: All About Jurats Notary Essentials: The Difference Between Acknowledgments And Jurats Additional Resources: NNA Hotline Email Share 37 Comments Add your commentCalvin C Wilson01 Jun 2015thank you very goodpatti01 Jun 2015top of screen is blank. have no idea what this isGary Halvorsen01 Jun 2015When notarizing a 4 page document (for example), is it legal to also put your stamp on the back side of all 4 pages (spaced out a little so the stamp is shown a little bit on each page). This then helps to ensure no additional pages have been added, or a page has been removed. When I was st sea I did this to cargo manifests and documents.National Notary Association04 Jun 2015Hello. To help ensure we provide you with the correct information for your state, can you please tell us what state you are commissioned in?Doylene Plank01 Jun 2015This is great information. I am keeping in my file for future reference.Regina Stevenson01 Jun 2015Can I return this course for a refundNational Notary Association02 Jun 2015Hi Regina. If you have questions regarding a refund for an NNA course or product, please contact our Customer Care team at 1-800-876-6827 or email@example.com and they can help you.Greg02 Jun 2015Very good information, and very timely. There is a civil litigation case currently ongoing in Seminole County, Florida in which the court declined to rule on judgment due to the fact that the plaintiff did not take, and the notary did not administer, an oath while executing an affidavit central to the case. A very real consequence of not having the oath/affirmation properly administered.Memo Reyes02 Jun 2015So when you're doing a Jurat, do you have ask the client if they want to take an Oaths or an Affirmations?National Notary Association04 Jun 2015Hello Memo. Yes, you should ask the signer if they have a preference between an oath or affirmation and let the signer choose which they firstname.lastname@example.com Jun 2015I just became a Notary in the state of Maryland. Are affirmations and/or oaths only required for Affidavit's?National Notary Association09 Jun 2015Hello. No, Notaries in Maryland are also authorized to administer oaths and affirmations for other situations that require a person to make a promise to tell the truth, such as a person giving oral testimony for a deposition.Rebecca15 Apr 2016California- I've been asked to perform an oath on paperwork on a divorce decree from another country. I don't think I can perform Oaths on alternate paperwork. Do I attach a jurat?National Notary Association15 Apr 2016Hello Rebecca. This article has some information that may be helpful to you: https://www.nationalnotary.org/notary-bulletin/blog/2015/07/notarizing-documents-from-other-countries. The signer must tell you what type of notarization they want-if unsure, the signer may need to contact the document's issuing agency for instructions. John Axt23 May 2016It may be simpler to look at this in religious terms. If a person does not believe in god they can use an affirmation instead of swearing to a god they don't believe in. In the practical world that notaries live in, you don't ask if they want an oath or affirmation. If a person objects to swearing an oath administer an affirmation. Just that simple.James23 May 2016Luckily, here in NJ, there is a law that specifically states that the "oath ceremony" that is, the uplifting of the hand and asking the signer to confirm "yes" or "no" isn't necessary to the validity of an oath or affirmation. However, I always like to ask because it gives them a sense of, "This is a serious thing. I should tell the truth here."Joan23 May 2016Can anyone suggest language for dealing with the "mhmm" response? Do you advise before hand that now you will now administer the oath/affirmation and need a clear verbal answer in response?Bernie Swanson24 May 2016I would include the words "Please respond with a verbal yes or no" after asking do you swear or affirm.....Anna Marie Mastrangelo23 Aug 2016I am a NSA, most of my notorrial acts are with in the context of a loan closing. There are up to 20 different documents within the packet that need to be notarized. When entering the info into my journal is it necessary to note each and every document or is one entry enough for the closing?National Notary Association23 Aug 2016Hello. To help us answer your question, can you please tell us what state you are commissioned in?Glenda Rooney 29 Mar 2017I love this information from NNA 😎Kathleen15 May 2017As usual, this information is very helpful - thank you!Jerry Lucas16 Apr 2018I wrote a blog article on the History of the Oath. The affirmation was created to accommodate Quakers, Mennonites and Moravians, who were religious, but believed that passages in the Bible prohibited taking an oath. In the past, oaths were also taken on sacred objects including stones, weapons, and even bear heads by Siberian hunters. Ancient oaths sometimes included a curse as a penalty for making false statements. See http://abclegaldocs.com/blog-Colorado-Notary/history-of-the-oath-930-ad/Kerri16 Apr 2018I'm a little confused in the course it says you can not notarize any kind of marriage certificate but doesn't say about an application for a marriage licenceKarrie30 Apr 2018When is an oath or affirmation necessary? Will he paper I'm notarizing say if we need to perform this? Is it only to be done when requested by a customer? I am a notary in Greene County, Missouri. Thank you!National Notary Association30 Apr 2018Hello. The Notary cannot choose what kind of notarization is needed-if the signer wishes to take an oath or affirmation, the signer must tell you that is the type of notarization they want. If the signer is unsure, the signer may need to contact the agency that issued the document to ask if an oath or affirmation is required. Please see this article for more information: https://www.nationalnotary.org/notary-bulletin/blog/2015/04/key-differences-acknowledgment-jurat-certificatesCheryl Cook07 May 2018When completing a Jurat and Oath or Affirmation, do I need two separate journal entries or can I just include everything on the one Journal entry?National Notary Association07 May 2018Hello. If you are administering the oath or affirmation as part of a jurat notarization, you only need one journal entry. James08 May 2018Here in NJ, the law states that the uplifting of the hand is not required when administering the oath as part of a jurat. The only requirement is that the person who is under oath, knows they are under oath, and that they consent to it (if not in open court) and that they understand that lying while under oath is perjury.Jaqueline M Perkins30 Oct 2018I need a notary at 1pm tomorrow to swear me in over the phone over a contempt of court and modification of alimony. All I need is 40 second s to a minute of your time. Please help me!!!Krisie Stocker03 Jun 2019When recording a Jurat and oath or affirmation in our journal, does the oath or affirmation require its own line entry?National Notary Association04 Jun 2019Hello. If the oath or affirmation was administered as part of the jurat process, then no. However, if the oath or affirmation was administered verbally as a separate notarization apart from the jurat, then it would require its own journal entry. Sherry Hendershot21 Aug 2019My boyfriend has to participate in a phone hearing out of state and needs a notary to administer an oath. I am a notary, can I administer the oath as his girlfriend?National Notary Association22 Aug 2019Hello. To help us answer your question, can you please tell us what state you are commissioned in?rosalie carrillo14 Oct 2019Just became a Notary in California... How much is the usual fee if any to administer and Oath or Affirmation?National Notary Association14 Oct 2019Hello. The maximum fee you may charge for a jurat, or for administering an oath or affirmation apart from a jurat is $15 per person (GC 8211 [a] and [b]).James14 Apr 2020Rule No. 1 flew right out the window with the national acceptance of Remote Notarizarions. What that tells me is that if you let that rule dissolve, why even worry about the rest of it.Leave a Comment Required * Name * Email *(for verfication purposes only) Comment * Enter the text shown in this image *(text is case sensitive)All comments are reviewed and if approved, will display.


text: Get In Touch With UsCONTACT US FORMName* Email Address* Phone* Are you a new patient? YesNoWhat services are you interested in? * Check all that apply ChiropracticStop Smoking ProgramLaser Weight Loss ProgramBraincore TherapyLaser Allergy ReliefAppointment DateAppointment DateAMPMMessage [recaptcha]Call The Office(914) 738 – 4460Office Location604 Fifth Avenue, Pelham NY 10803Email Us[email protected]Click on map for directions

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Mahala Candle
Presented in a beautiful designer gift box, the Mahala Candle is available in six beautiful and natural fragrances derived from the essence of fruits, plants and trees. Collect all six and treat yourself on those cold winter days, lazing on the couch huddled under our Korbel Blanket with hot chocolate and a good book or have on standby for a special gift idea.
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Monday, October 4, 2010
better late than never.....
Grab This Button
Hey readers! this is the first year I get to participate in 31 for 21 as the proud parent of a child with Down Syndrome! Im not quite sure how I am going to "do this" but if you know me I will probably just jump right in with both feet. I am also going to snoop around and see if I can find a few more blogs that have some information that could be useful as we navigate through the ups and downs of it all......
This weekend was FULL!
My parents came down to meet Miss G.
I hadn't put it on my calendar because sometimes life throws a curve ball and I didn't want to get my hopes up, only to be disappointed that they changed their mind or to find out that circumstances helped them. Since Mom and Dad drove down my sister and her family came too. They all stayed with my Grandpa and that meant a FULL house for him. My cousin and her daughter are staying with him, then my aunts and uncles that live close by drove over each day and we just enjoyed hanging out together. The weather was incredible!!!! (today I am actually COLD!) Last night my Sisters family and parents came our way to have supper. Paul cooked a new "Alton Brown recipe" ..... AWESOME pork tenderloin with lime and chipotle pepper marinade. Listen ladies, you HAVE to give this a try. I cant wait to do it with brown rice as a side to drizzle the marinade/ juice on. It was easy, or at least Paul said it was.... ;) Yes I thanked him. I made sweet potatoes, weve been enjoying them peeled and tossed with olive oil salt and baked in the oven.... YUM-O. My daughter and her husband and my son and his girlfriend were here. (Ive asked them to save Sunday evenings for me.) I really feel the need to have all my peeps in the nest once a week at least. I know they are busy, I understand when things come up, but to have a goal is good.Donchathink???
I enjoyed watching Miss G interact with the family, her reaction to the noise and business.... some moments were better than others. Ive decided that I am going to call my cousin and have play dates with her daughter. They will be here until December I think. It will be good for Miss G to learn some playtime etiquette and for us to be with my grandpa. He doesnt live that far away but I just have a hard time getting over to visit because its out of the way.
I also wanted to tell you that G has been asking to watch her signing time DVDs. This morning she signed " signing time, help" and stood there waiting with those big brown eyes for me to get up and put her DVD on. I have been visiting the website and I am trying to pick out the next video to purchase.
It would be logical to pick number 4 since we have 1-3
But if you know me,
I dont always do whats "expected".....
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Utilizing credit to cover life's major expenses or unexpected bills is common among Americans. Taking on debt makes it easier to manage a significant financial burden when the savings just isn't there. Unfortunately, leaning too heavily on credit can create a mountain of debt that feels impossible to move.
If you feel like you're drowning under its weight, you are far from alone. A recent study from LendingTree highlights the glaring truth of debt across the United States – consumers are spending 26% of disposable income on non-housing debt in the form of credit card balances, auto or personal loans, and medical debt. As these financial obligations roll over from month to month, interest accumulates, often in the double digits, making it seem like a never-ending cycle of paying someone else.
When you're in debt, you may think you're drowning financially, but the truth is, there is a way out. You may not be able to see that path forward just yet, but simple steps can get you back on track toward a debt-free life. It begins with having a plan to climb out from under your debt burden with the following tips and tools in mind.
First, Know Where You Stand
It isn't feasible to create a plan to get out of debt if you don't first know where you stand. Many people take the convenient path of avoiding their debt and simply paying the minimums due each month to stay current. However, that doesn't make a dent in the balance owed, especially when interest is accruing every month. Instead of ignoring the issue, you have to take a hard look at what you owe, including your current balances, the interest rate on each debt, and monthly payment due.
With this information gathered, you can most effectively determine your course of action for getting out from under your debt load. Seeing all the numbers laid out in front of you may be overwhelming at first, but once you have the big picture, you can start to see a light at the end of the tunnel.
Check Your Budget – and Your Credit
After gathering all the details about the debts you owe, your next step is to evaluate your cash flow. It is easy to know what comes in each month in terms of income, but where most people struggle is the outflow. Understanding your expenses in both the required and desired categories is essential to getting out of debt.
Required expenses are easy to calculate – think of your housing payment, debt obligations (the minimums), utilities, and other necessary bills each month. Desired expenses include anything not on the first list. For most, these costs are dining out and entertainment, impulse buys, and other non-essential items. Add the two categories up and subtract the number from your income to see how much wiggle room you have each month, if any. When a surplus is available, plan to use that toward speeding up your debt pay off. If you have a deficit or are just breaking even, consider what you can cut out of that desired category to free up some extra cash.
It is also important to check your credit. When you feel like you are drowning in debt, pulling your credit report may seem like an unnecessary and depressing task. However, checking your credit gives you an opportunity to see where you stand as well as recognize any errors that may be on your report. When errors are present, work toward disputing those items to help clean up your credit score. This will help you improve your overall financial standing with new creditors, making it easier to get more affordable access to credit in the future. You can check your credit with each of the three credit bureaus for free online.
Recognize Problem Areas
One of the most challenging aspects of getting out of debt is not necessarily tied to minimal income but instead, it links back to behaviors with money. If you really want to stop drowning in debt, you need to evaluate how you get into the situation in the first place. Maybe you're desired spending was above and beyond your means, or you have a fondness for online shopping. Whatever the issue may be, consider how you can correct it or at least reign it in moving forward. Getting out of debt requires not adding to your balances each month; that means putting an end to unhealthy spending habits now.
Pick a Strategy
Once you have done to work to evaluate your debt situation, your cash flow, and your spending habits, it's time to consider your options for getting out of debt. There are a handful of strategies you can use to effectively pay off your debts, including the snowball method and the avalanche method. The first, the snowball method, means you start paying off your smallest debt first to build momentum. Extra funds you have each month go toward this balance, while you pay the minimum on the others. The momentum that comes with paying off the first balance trickles down to the next, and the next, until you have paid off your debts in full.
The avalanche method works differently. Instead of focusing on the smallest balance, you put your extra cash toward the debt with the highest interest rate. Payment on other debts is still the minimum amount, but once you have paid off the most costly obligation, you transfer that amount to the next-highest-rate debt. Doing so helps save you money in the long run, as you are accumulating less interest on your debts over time. Either method can work, but both require understanding the details of your debt situation first.
Have a Plan for the Future
Feeling like you are drowning in debt can be an awful feeling, but you have a way out if you choose to put in the work. In creating a plan to pay off what you owe, consider what you want for the future as well. Being debt-free is often the most pressing objective, but you can easily get off track if there is not a bigger-picture goal behind it.
For instance, think about what you could do with the money no longer being used toward debt payments. How much could you save with those funds over time? Could you build up retirement savings or invest for your longer-term goals? Answering these financial questions is part of creating a strategic plan for getting out from under the weight of debt, and being successful in doing so.


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All shy guys are faced with one dilemma – to let know or to not let know – when it comes to their love interest. Usually , they never let you know directly unless you give some body language. Here are some signs that a shy guy likes you.
1. He Avoids Direct Eye Contact:
If a shy guy is in love with you, he would talk through his eyes but not directly. Shy guys stare if they like you, that's a fact, but they are quick to avoid your eye contact if you gaze in their direction. They just can't maintain eye contact for long and they will either look down or sideways. He would stare almost unconsciously at you but when he sees you looking at him he will become very self conscious and avert his gaze. He might give you a polite smile if you catch him staring but he will look away while doing so.
2. He Blushes When You Talk to Him:
You can see a pink flush on his cheeks if you flirt with him or get close to him or even if you just have a casual conversation. Shy guys are overly sensitive and self conscious, he's been thinking or even obsessing about you in his mind and when you talk to him, he blushes.
3. He Becomes Self Conscious When You are Around:
If you see him touching up his hair, scratching his face, twirling his fingers or getting fidgety whenever you are around, it would indicate that he has become very self conscious in your presence. This is definite sign of a shy guy liking you.
4. He Stammers While Talking to You:
How to know if a shy guy likes you? Just talk to him and you will know. Just start a casual conversation and see how he reacts to you. If he stammers back a reply, while looking all around the room but at you, then you know he really likes you.
5. He Gets Your Number from Your Friends and Calls You Up:
It is quite natural for a shy guy to never ask for your number directly. If he's really interested in you, he might take up the courage to get your number from your friends, and then call you up.
6. He is Very Sensitive to Your Touch:
If you happen to touch him playfully, or just casually, he might respond as if he's got an electric shock. You will see his face go all pink, he might suddenly move away or just get overly self-conscious. It just shows your touch means a lot to him.
7.He Offers to Help You With Your Work:
This might be one way he will try to approach you. He will offer to help you with your work or offer to drop you home. This is a big step for a shy guy and you should take it as a definite sign that he likes you and is in love with you.


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The biggest issue facing Connecticut is not the economy -- it's denial. Denial that this state can survive another Democratic administration.
The recent midterm election further cemented that Connecticut is in clear denial of the past failed Democratic polices. On Tuesday, the state decided to double-down and expand the Democratic majorities at the state and federal level, a mind-blowing achievement when you consider the state is insolvent, unfunded liabilities are soaring and corporations and families are departing for more attractive states.
And the state decided to elect more of the same. The recent election results will only accelerate the migration of talent and taxes already departing in droves.
According to US News & World Report states ranking, Connecticut ranks at the bottom with respect to taxes, economy, infrastructure and cost of living. It is important to note that New Hampshire and Massachusetts both ranked overall in the top 10.
Connecticut is clearly in financial disarray with no clear path to prosperity. Governor-elect Ned Lamont has proposed tolls, moderate property tax adjustments and a willingness to reform the state pension dilemma. His ideas clearly resonated with the state electorate, but I suspect as always with this state it will prove to be nothing more than words.
Tuesday was not a good day for this state. We buried our heads further in the sand and elected the same philosophy that got us here.
David Johnson, Simsbury


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Osteoclasts are the principal bone-resorbing cell in the body. An ability to isolate osteoclasts in large numbers has resulted in significant advances in the understanding of osteoclast biology. In this protocol, we describe a method for isolation, cultivating and quantifying osteoclast activity in vitro.
Osteoclasts are highly specialized cells that are derived from the monocyte/macrophage lineage of the bone marrow. Their unique ability to resorb both the organic and inorganic matrices of bone means that they play a key role in regulating skeletal remodeling. Together, osteoblasts and osteoclasts are responsible for the dynamic coupling process that involves both bone resorption and bone formation acting together to maintain the normal skeleton during health and disease.
As the principal bone-resorbing cell in the body, changes in osteoclast differentiation or function can result in profound effects in the body. Diseases associated with altered osteoclast function can range in severity from lethal neonatal disease due to failure to form a marrow space for hematopoiesis, to more commonly observed pathologies such as osteoporosis, in which excessive osteoclastic bone resorption predisposes to fracture formation.
An ability to isolate osteoclasts in high numbers in vitro has allowed for significant advances in the understanding of the bone remodeling cycle and has paved the way for the discovery of novel therapeutic strategies that combat these diseases.
Here, we describe a protocol to isolate and cultivate osteoclasts from mouse bone marrow that will yield large numbers of osteoclasts.
Bone remodeling is dynamic and involves the coupling of bone formation with bone resorption1. This tightly regulated process is responsible for maintaining the skeleton during normal homeostasis, and in response to injury and disease.
Osteoclasts are unique, multinucleated cells that are capable of resorbing both the organic and inorganic matrices of bone. Osteoclasts are derived from the monocyte/macrophage lineage of the bone marrow2-5. Abnormalities in the function or formation of osteoclasts can result in a variety of clinical pathologies, including common conditions like osteoporosis.
The ability to generate osteoclasts in vitro has allowed for significant advances in our understanding of bone biology6. As a result, new therapeutic agents are emerging to treat osteoclast-related diseases which are responsible for significant morbidities and mortalities7. Homeostatic maintenance of bone mass and strength requires the concerted action of bone-forming osteoblasts and bone-resorbing osteoclasts8,9. Bone homeostasis is altered in a number of diseases, including post-menopausal osteoporosis, in which increased osteoclast activity leads to pathogenic loss of bone mass and density10. With increasing availability of transgenic murine models of human disease, there is more opportunity to decipher the role of the osteoclasts in human bone disease11-13.
Numerous protocols for osteoclast culturing techniques appear in the literature, with many variations described9,12,14. Xing and colleagues describe similar methodology to the protocol described below, in their description of osteoclastogenic assays from murine bone marrow cells. However to release the bone marrow cells following long bone harvest, Xing et al. flush the marrow cavity with α-MEM complete media14. Catalfamo examines the effect of hyperglycemia on osteoclast function and describes a method in which all cells mobilized by bone marrow flushing are cultured for 24 hr, at which point the non-adherent cells are discarded12, a technique also used by Boyle et al.9 These previously published protocols necessitate the practice of flushing the bone marrow, a tedious practice, which also introduces the risk of a needle stick injury and loss of valuable bone marrow, as one must cut both ends of the bone. The protocol, which we describe, implements the use of a mortar and pestle to isolate osteoclasts, which is similar to the method of macrophage isolation described by Weischenfeldt et al.15
Our experience, however, is that osteoclast isolation and in vitro culture using previously published techniques results in variable outcomes in terms of osteoclast production, often resulting in an inability to cultivate osteoclasts. Therefore, we have devised a protocol that allows for the consistent isolation of mouse bone marrow to produce large numbers of multinucleated osteoclasts in vitro, with an approximate yield of 70-80% of cells initially plated forming macrophages and subsequently osteoclasts, in the presence of osteoclast induction media.
Subscription Required. Please recommend JoVE to your librarian.
NOTE: Ethical statement: All research involving vertebrate animals was performed in accordance protocols approved by the Stanford Administrative Panel on Laboratory Animal Care (APLAC).
1. Preparation
- Allow 10 ml of commercially available density gradient cell separation media (which contains polysucrose and sodium diatrizoate, adjusted to a density of 1.077 g/ml) to come to RT in a 50 ml conical tube.
- Prepare flow cytometry (FACS) buffer with 1x phosphate buffered saline (PBS) and 2% fetal calf serum (FCS). Take a 50 ml aliquot and maintain this aliquot at RT for use during the density gradient cell separation media step.
- Have a bucket of ice ready to maintain isolated tissue and cells in during the isolation procedure.
2. Prepare Culture Media
- Prepare basal media: MEM (Minimal Essential Medium), without phenol red (500 ml), glutamate (1x), FCS (10x), 10,000 units/ml penicillin (1x).
- Filter basal media with a 0.22 μm filter.
- Prepare bone marrow macrophage induction media:
- Take 50 ml of the basal media prepared in Step 2.1.
- Add Prostaglandin E2 (PGE2, Mr 352.465 g/mol) at 10-7 M final concentration.
- Add Macrophage-Colony Stimulating Factor (M-CSF) at 10 ng/ml.
- Do not filter the final solution of bone marrow macrophage stimulating media.
- Prepare osteoclast induction media:
- Take 50 ml of the basal media prepared in Step 2.1.
- Add Prostaglandin E2 (PGE2, Mr 352.465 g/mol) at 10-7 M final concentration
- Add Macrophage-Colony Stimulating Factor (M-CSF) at 10 ng/ml.
- Add RANKL at 10 ng/ml.
- Do not filter the final solution of osteoclast induction media.
3. Bone Marrow Isolation
- Euthanize one C57BL/6 mouse using carbon dioxide inhalation method, followed by cervical dislocation.
NOTE: Rodents must be euthanized by trained personnel using appropriate technique, equipment and agents.
- Position and secure the mouse on a dissection board and spray with 70% alcohol.
- Dissect out the femurs, tibiae, humeri and spine.
- Begin with the mouse in supine position. Make an incision vertically along the lower limb and begin by dissecting along the length of the femur and tibia by dividing soft tissue attachments. Finally, dislocate bones from the knee and hip joint to free bones fully from the skeleton.
- Proceed to make an incision along the length of the upper limb to isolate the humerus. Dislocate the humerus at the shoulder and elbow joints by dissecting soft tissue capsular attachment.
- Finally, turn the mouse over, so that the mouse lies prone. Make a skin incision along the length of the spine, in the midline. Observe the paravertebral soft tissue on each side of the spine and incise along the bony edge of the spine, dissecting through the paravertebral soft tissue mass. Using a scissors, make a horizontal cut across the spine at the base of the neck, and at the end of the spine, just proximal to the tail insertion, freeing the spine from soft tissue attachment.
- Once the bones are harvested, keep them in FACS buffer on ice.
- Use tissue paper to clean muscle and soft tissue from the bones.
- Place the cleaned bones into a pestle and mortar with 3 ml of FACS buffer.
- Crush the bones gently in the pestle and mortar.
- Aspirate off the blood-stained fluid and transfer into a 50 ml conical tube by passing the solution through a 70 μm cell strainer.
- Add 5 ml FACS buffer to the crushed bone and crush further. Again, remove the fluid using a pipette and transfer to the 50 ml conical tube through a 70 μm strainer.
- Typically crush the bones twice to three times in fresh FACS buffer, using a mortar and pestle, each time adding a further 5 ml of fresh FACS buffer, until the fluid does not stain red.
- Centrifuge the bone marrow solution at 200 x g for 5 min at 4 °C to yield a cell pellet.
4. Gradient Separation of Bone Marrow Cells
- Aspirate the supernatant and resuspend in 10 ml of FACS buffer (maintained at RT).
- Take the 50 ml conical tube containing 10 ml of commercially available density gradient cell separation media (RT).
- Layer the bone marrow solution onto the density gradient cell separation media. Do this slowly using an electric pipette. Angle the pipette tip against the inner surface near the top of the conical tube and tilt the conical tube to about 30°. Using a slow speed setting on the pipette, gently expel the bone marrow solution so as not to disturb the density gradient cell separation media.
NOTE: Ultimately there will be a clear definition between the two solutions (cellular solution and density gradient cell separation media solution).
- Using a RT centrifuge, centrifuge the density gradient cell separation media and cell solution in a balanced centrifuge at 200 x g for 15 min. In addition, remove acceleration and deceleration on the centrifuge to ensure adequate phase separation using the density gradient cell separation media gradient.
- After centrifugation, aspirate off the cloudy middle layer which contains the bone marrow cells of interest.
- Transfer these cells into a new conical tube. Add 20 ml of additional FACS buffer (kept on ice) to wash the cells.
- Centrifuge at 200 x g for 5 min at 4 °C to yield a cell pellet.
- Resuspend the final cell pellet in 1ml of macrophage stimulating media to allow for cell counting using a hemocytometer.
5. Cell Counting
- Take 10 µl of the cell solution and mix with 10 μl of trypan blue. Load 10 µl of the resultant mix onto a hemocytometer and obtain the cell count per ml of solution.
6. Cell Culturing
- Place 2 ml of macrophage stimulating media into each well of a 24-well plate.
- Add 200,000 cells to each well.
NOTE: In our experience, cell-plating density is one of the most crucial requirements for adequate in vitro culturing of osteoclasts.
- Gently agitate the plate and place in a regular incubator at 37 °C.
- Do not change the media for 3 days.
- After 3 days in macrophage stimulating media, change the media to osteoclast media.
- Hereafter, change the media daily for 5–7 days, at which point, large, multinucleated osteoclasts should be visible on the culture plate.
7. Staining with Tartrate Resistant Acid Phosphatase (TRAP)
- After 5–7 days of in vitro culture, aspirate the media from the culture well.
- Wash the wells gently with 1x PBS three times.
- Fix the cells by adding 4% paraformaldehyde solution and leave for 1 hr at 4 °C.
- During this time, prepare the TRAP stain (which is commercially available) by pre-warming to 37 °C.
- After 1 hr in paraformaldehyde, aspirate off paraformaldehyde and wash gently 3 times with 1x PBS.
- Add 1 ml of pre-warmed TRAP stain into each well of the plate to be stained.
- Place the plate in an incubator at 37 °C for 1 hr, shielded from light.
- After 1 hr, aspirate off the TRAP stain.
- Wash the wells 3 times with pre-warmed deionized water.
- Counterstain the well with Gill's Hematoxylin (CAUTION) for 1–2 min.
- Image osteoclasts using bright field microscopy.
- Wash wells with deionized water until an adequate color intensity of the stain is achieved, typically when nuclei appear blue.
8. Osteoclast Resorption Assay
- Use a commercially available 24-well polystyrene culture dish that is pre-coated with either bone substrate or an inorganic crystalline calcium phosphate coating.
- Place 2 ml of macrophage stimulating media into each well.
- Add 200,000 freshly isolated bone marrow cells obtained at the end of step 2 to each well.
- Gently agitate the plate prior to placing it in an incubator at 37 °C for 3 days.
- On day 3, change the media to osteoclast differentiation media.
- Perform daily media changes with osteoclast differentiation media for 5–7 days.
9. Quantification of Osteoclast Resorption Activity
- After 5–7 days of in vitro culture in osteoclast differentiation media on a mineralized substrate, analyze the mineralized surface, as described below, to assess the ability of osteoclasts to form resorption pits, an indicator of osteoclastic activity.
- Aspirate media from the resorption assay plate and add 100 μl of 10% bleach solution to each well.
- Incubate the cells with the 10% bleach solution for 15 min at RT.
- Wash the wells 3 times with distilled water.
- Aspirate off any remaining water and shake off excess water from the plate and allow to air dry for 3–5 hr.
- Counterstain the plate with a commercially available Von Kossa staining kit to allow for easier visualization of the un-resorbed substrate.
- Use bright field microscopy to take images of representative parts of the resorbed surface.
- Using image analysis software, calculate the percentage of the resorbed surface to allow for quantification of osteoclastic resorptive activity.
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Representative Results
The aim of this method was to easily isolate large numbers of osteoclasts in vitro, typically in one week. Successful isolation of large numbers of osteoclasts was confirmed using tartrate-resistant acid phosphatase staining (Figure 1A). Large osteoclasts are visualized as large purple cells with multiple nuclei (typically ≥ 3 nuclei). Using this protocol, it is common to isolate osteoclasts with as many as 30 nuclei per osteoclast (Figure 1B).
Using a mineralized surface is considered the gold standard method of assessing osteoclast resorptive activity in vitro. Using this method, the percentage of mineralized surface, or "resorption pits" that have formed allows for determination of osteoclast resorptive capacity (Figure 2).
Figure 1. (A) Tartrate resistant acid phosphatase staining of cultured bone marrow following 10 days in culture under osteoclast inductive conditions. Bright field micrograph at 10X magnification demonstrates multiple large, multinucleated osteoclasts that are TRAP positive (purple staining). An example of a large, multinucleated osteoclast is outlined by a red dashed line. (B) Bright field micrograph at 10X magnification demonstrating a large, multinucleated, TRAP positive osteoclast. An example of a nucleus within a multinucleated osteoclast is shown by the red arrow. Please click here to view a larger version of this figure.
Figure 2. Bright field micrograph at 10X magnification demonstrates resorption pits formed by osteoclastic resorption activity. Mineralized matrix is stained with Von Kossa stain to allow for visualization. The black dashed lines outline an example of resorbed areas of matrix. Please click here to view a larger version of this figure.
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An ability to easily isolate and cultivate large numbers of osteoclasts in vitro has been responsible for helping to advance understanding of bone biology and osteoclast-mediated diseases. It was the identification of RANKL that lead to this, when it was recently identified as the major regulator of osteoclast formation, differentiation and survival16-18.
It has been our experience that the in vitro cultivation of osteoclasts from bone marrow is largely dependent on seeding density. We observed that failure of this protocol to yield osteoclasts is typically due to a suboptimal seeding density when plating bone marrow-derived cells. Thus, in this protocol, it is recommended to seed 200,000 cells per 24-well plate well. Furthermore, in order to achieve maximum success with this technique, it is recommended to prepare the osteoclast induction media precisely as described above and to change the media at the same time each day, after the initial 3 days when one does not need to disturb the culture plates. This may allow a constant concentration of the osteoclast induction media factors. In addition, we have found that the use of specific reagents also allows greater potential for culturing of large numbers of multinucleated osteoclasts.
This technique is limited by the availability of a RT centrifuge, in which one can alter acceleration and deceleration parameters and procurement of specific reagents. It is otherwise a very straightforward protocol, with which we have had much success, in a range of mice of differing age and genetic background (e.g., C57BL/6, CD1, NOD SCID, Lep db-/-).
This technique offers a reliable method to generate osteoclasts in vitro and to determine their ability to resorb mineralized matrix. Using this protocol, it is possible to isolate large numbers of multinucleated osteoclasts in vitro typically in one week. Our experience is that osteoclast isolation and in vitro culture using previously published techniques results in highly variable outcomes in terms of osteoclast production, often resulting in an inability to cultivate osteoclasts9,12,14.
Osteoclast function is complex and bone resorption is a highly orchestrated process that is mediated at the local level by cross-talk between osteoblasts and osteoclasts19. In addition to local control of bone balance, emerging evidence suggests that osteoclast activity is governed systemically by factors related to immune, neuronal, and other systematic factors20-24.
As the understanding of osteoclast biology continues to advance rapidly, including their diverse role in processes ranging from bone remodeling to their role in regulating cancer metastases to bone25-27, this protocol will allow researchers to consistently isolate large quantities of osteoclasts in vitro.
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None of the authors has disclosure or conflict of interest to declare.
We acknowledge the support of NIH grants R01 DE021683, R01 DE019434, U01 HL099776, The Oak Foundation and The Hagey Laboratory for Pediatric Regenerative Medicine.
|Catalog Number
|MEM, no glutamine, no phenol red
|M-CSF, recombinant mouse
|Recombinant Mouse TRANCE/RANK L/TNFSF11 (E. coli expressed)
|R&D Systems
|Prostaglandin E2
|Acid Phosphatase, Lekocyte (TRAP) kit
|Osteoassay bone resorption plates, 24 well plates
|Corning Life Sciences
- Sims, N. A., Martin, T. J. Coupling the activities of bone formation and resorption: a multitude of signals within the basic multicellular unit. BoneKEy reports. 3, 481 (2014).
- Kahn, A. J., Simmons, D. J. Investigation of cell lineage in bone using a chimaera of chick and quial embryonic tissue. Nature. 258, 325-327 (1975).
- Walker, D. G. Bone resorption restored in osteopetrotic mice by transplants of normal bone marrow and spleen cells. Science. 190, 784-785 (1975).
- Burger, E. H., et al. In vitro formation of osteoclasts from long-term cultures of bone marrow mononuclear phagocytes. The Journal of experimental medicine. 156, 1604-1614 (1982).
- Underwood, J. C. From where comes the osteoclast. The Journal of pathology. 144, 225-226 (1984).
- Lacey, D. L., et al. Bench to bedside: elucidation of the OPG-RANK-RANKL pathway and the development of denosumab. Nature reviews. Drug discovery. 11, 401-419 (2012).
- Brown, J. E., Coleman, R. E. Denosumab in patients with cancer-a surgical strike against the osteoclast. Nature reviews. Clinical oncology. 9, 110-118 (2012).
- Khosla, S. Minireview: the OPG/RANKL/RANK system. Endocrinology. 142, 5050-5055 (2001).
- Boyle, D. L., et al. Differential roles of MAPK kinases MKK3 and MKK6 in osteoclastogenesis and bone loss. PloS one. 9, (2014).
- Hofbauer, L. C., Heufelder, A. E. Role of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in bone cell biology. Journal of molecular medicine (Berlin, Germany). 79, 243-253 (2001).
- Teramachi, J., et al. Increased IL-6 Expression in Osteoclasts is Necessary but not Sufficient for the Development of Paget's Disease of Bone. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research. (2013).
- Catalfamo, D. L., et al. Hyperglycemia induced and intrinsic alterations in type 2 diabetes-derived osteoclast function. Oral diseases. 19, 303-312 (2013).
- Schueler, J., et al. Intratibial injection of human multiple myeloma cells in NOD/SCID IL-2Rgamma(null) mice mimics human myeloma and serves as a valuable tool for the development of anticancer strategies. PloS one. 8, (2013).
- Xing, L., Boyce, B. F. RANKL-Based Osteoclastogenic Assays from Murine Bone Marrow Cells. Methods in molecular biology (Clifton, N.J). 1130, 307-313 (2014).
- Weischenfeldt, J., Porse, B. Bone Marrow-Derived Macrophages (BMM): Isolation and Applications. CSH protocols. (2008).
- Yamamoto, Y., et al. Osteoblasts provide a suitable microenvironment for the action of receptor activator of nuclear factor-kappaB ligand. Endocrinology. 147, 3366-3374 (2006).
- Yasuda, H., et al. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proceedings of the National Academy of Sciences of the United States of America. 95, 3597-3602 (1998).
- Lacey, D. L., et al. Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 93, 165-176 (1998).
- Teitelbaum, S. L., Ross, F. P. Genetic regulation of osteoclast development and function. Nature reviews. Genetics. 4, 638-649 (2003).
- Agas, D., Sabbieti, M. G., Marchetti, L. Endocrine disruptors and bone metabolism. Archives of toxicology. 87, 735-751 (2013).
- Manolagas, S. C., O'Brien, C. A., Almeida, M. The role of estrogen and androgen receptors in bone health and disease. Nature Reviews Endocrinology. 9, 699-712 (2013).
- Martin, T. J., Udagawa, N. Hormonal regulation of osteoclast function. Trends in endocrinology and metabolism. 9, 6-12 (1998).
- Nakamura, T., et al. Estrogen prevents bone loss via estrogen receptor alpha and induction of Fas ligand in osteoclasts. Cell. 130, 811-823 (2007).
- Bellido, T., et al. Regulation of interleukin-6, osteoclastogenesis, and bone mass by androgens. The role of the androgen receptor. The Journal of clinical investigation. 95, 2886-2895 (1995).
- Roato, I. Interaction among cells of bone, immune system, and solid tumors leads to bone metastases. Clinica., & developmental immunology. 2013, (2013).
- Autio, K. A., Morris, M. J. Targeting bone physiology for the treatment of metastatic prostate cancer. Clinical advances in hematolog., & oncology. 11, 134-143 (2013).
- Sottnik, J. L., Keller, E. T. Understanding and targeting osteoclastic activity in prostate cancer bone metastases. Current molecular medicine. 13, 626-639 (2013).


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I got my new passport today. I'm good to travel for another ten years.
I wanted to move my Indefinite Leave to Remain visa from my old passport to the new one. Seems like it would be a fairly straightforward procedure. Given everything I've paid in fees in the past, you would think it is would be free. Okay, maybe there would be a small administrative charge for the transferring the sticker, or even pasting a new one in the new passport.
Not exactly. There is a £160 fee. It's like a 10-year recurring tax to be a taxpayer. But that's for having it done by post. So I'll just take in personally and have it done. Less administrative hassle for the bureaucrats, so a much cheaper fee, right?
Not exactly. The fee goes up. Way up. So what does the Government charge me for using my own petrol and taking time off work to make things easier for them? £500. There is an advantage to me though. I don't have to wait up to 14 weeks to get my documents returned. So I suppose I'm paying for the privilege of not being prevented from traveling for three months.
But there's more. Just as if I was applying to enter the UK for the first time, I have to answer the usual questions:
In times of either peace or war have you or any dependants included in this application ever been involved, or suspected of involvement, in war crimes, crimes against humanity or genocide?
Have you or any dependants included in this application ever been involved in, supported or encouraged terrorist activities in any country?
Have you or any dependants included in this application ever been a member of, or given support to, an organisation which has been concerned in terrorism?
Why do they ask these questions? Do they honestly think that someone is going to be involved in genocide or terrorism and then admit to it on a government form? This is honestly sillier that then question at the airline check-in counter about whether you packed your own bag, as if you are suddenly going to remember that a strange Middle Eastern man showed up at your house and asked if he could pack your bag for you, just as a random act of kindness.
Fortunately, I don't have to answer the questions (which wouldn't be a problem) or pay the £160 (which would), as long as I keep my old passport with me. I just have to present both documents when I want to get back into the country. As I see it, why should I pay £160 when it is going to cost me £655 to apply for citizenship and it will take the same amount of time to process the application?
It is much cheaper to become an American citizen. $330 (or about £165). A replacement green card is $190 (£95). Is this just another example of Rip-off Britain?


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American Politics Today (Third Essentials Edition)
All the tools students need to understand American politics—in a briefer text. American Politics Today helps students understand the debates and controversies that they encounter in the news by emphasizing conflict and compromise as natural parts of politics. The Essentials Edition includes all the learning tools from the full book—including highly visual "How It Works"
(as of April 20, 2020 6:12 am GMT+0000 - Details) + Add To Wish List


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Have you started feeling that year-end tingle of excitement running through your office yet? A colleague mentions a holiday plan or you get a glimpse of blue sky through the window, and you know…you're almost there! If your office is lagging on the final-stretch feelings, it may be because no one has started planning the year-end function. Someone needs to get on that, right? Yes!
Year-end functions are more than parties or frivolous distractions. They provide a little injection of excitement to the last quarter that may just be what your workplace needs to end the year on a high note. Let's be honest: it's almost November, and not only has everyone been working hard in the office, there have probably been personal challenges, setbacks and lots of ups and downs. Why not boost morale right now by starting to plan your year-end event? You'd be surprised how quickly people perk up at the thought of a fun social occasion on the horizon.
Not sure of how to make your year-end function special? Wondering about appropriate entertainment or budget friendly enhancements? A QuikPix Photo Booth at your year-end function takes care of all those concerns. Not only do your guests have a lot of fun on arrival, but they also get to take the memories home with them. We love seeing guests place photo booth strips on their desks back at the office as a reminder of a great night, and a great end to a beautiful year!
Interested in hiring a photo booth for your year-end function? See photo booth hire Johannesburg for more information.


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Earn points toward FREE PIZZA with Piece of the Pie Rewards
More About Domino's Near Harriman
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Get started by selecting your favorite crust. Choose from a hand-tossed crust that's overflowing with buttery flavor, a crispy thin crust, a pan crust featuring two delicious layers of cheese, or even a gluten-free pizza crust! Then, choose a sauce and pile on as many or as few pizza toppings as you want. Whether you're all about the veggies like green peppers, spinach, and onions, or you prefer the sweet and savory combination of ham and pineapple, you can make your dreams come true at your nearest Domino's!
With millions of different ways to craft a single Domino's pizza (yes, we said millions and we mean it), you can be sure we have a slice that suits your tastes. From its sauce to its size, build your own pizza or choose from one of our famous specialty pizzas, like the ever popular Buffalo Chicken, Pacific Veggie, or Ultimate Pepperoni and have it delivered hot and fresh to your doorstep, fast!
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Shot in Barcelona, starring Sophia Ndaba.
Thumbnail by Adrien Crasnault.
Shot in Barcelona, starring Sophia Ndaba.
Thumbnail by Adrien Crasnault.
Instagram @gregmirzoyan
December 23, 2016 in glam, photography
Tagged barcelona, bcn, blond, body, foto, girl, glam, mujer, photo, photography, see thru, sexy, woman
Posted in glam, greg mirzoyan, mamiya, photography
Tagged 120mm, 120mm film, beautiful, beauty, diapo, foto, fotografo, glamour, greg mirzoyan, mamiya, Mamiya RZ67 Pro II, medium format, medium format camera, mujer, photographer, photography, slide film, topless, woman
Posted in black and white, glam, greg mirzoyan, mamiya, photography, polaroid
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February 26 2013
D-Now Session Four: When God Says Wait
Ryan Jasper continues his D-Now series on prayer
Ryan Jasper continues his D-Now series on prayer
This entry was posted on Tuesday, February 26th, 2013 at 12:06 pm and is filed under , , Sermon. You can follow any responses to this entry through the RSS 2.0 feed. You can leave a response , or trackback from your own site.
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Welcome to the official blog and media hub for The HANGAR | Gallery & Arts Initiative. Located in the Wynwood Arts District, The Hangar is dedicated to the collective movement of creative exhibition and seeks to open the lines of communication between artist, patron and community. www.hangargallery.com
I'm Not From Around Here…
A Jacked Fashion Photo Shoot
Be sure not to miss the alien editorial coming soon!


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It took me a while to get into Penny Dreadful when it first aired in the UK. What I thought was going to be a slow and plodding historical horror took a few unexpected twists and turns and after a couple of episodes had me hooked. My one main complaint of the series was that it seemed to end too soon, a couple more episodes would have tied it all up quite nicely whereas I felt a sense of unfinished business the way they left it.
Anyway, my gripes aside it looks like they are making a sequel and this will move from late Victorian London to 1930's cosmopolitan Los Angeles.
I'm looking forward to it but I can't help thinking the producers have missed a trick and that setting the series in Chicago at the World Fair in the 1890's would have been the way to go. It would have allowed crossover from characters we already know and we could have checked out H.H Holmes and his Murder Castle!


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Be sure today to stop over at the Bobunny Blog- we are having a super fun day showing off the GORGEOUS new releases, and a fun scavenger hunt game- to win prizes! Make sure you also hop to the team blogs, as many of the girls have talked about giving away Blog Candy!! OHHH!! doesn't that sound fab?
thanks for coming over!


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Kyrgyzstan is unbelievably beautiful. Almost every place I've been here has a backdrop of dark, craggy mountains still covered with snow. We've been through dry, scrubby desert; alpine meadows; red canyons; high mountain passes where snow by the road still hasn't melted.
I spent two nights sleeping in a yurt in a high (10,000 feet above sea level) meadow, green and smooth and grassy, where sheep and cows and compact Kyrgyz horses graze. It's just above Song-Kul Lake, which – like all lakes in this country, it seems – is a deep blue, even on a cloudy day. The valley lies up a rough track off the main dirt road, which itself follows the lake and cuts between two snow-covered mountain ranges. As we drove between the lake and the mountains, we saw a gathering of horses and people and cars, so we stopped the truck and walked up the hill to investigate.
It was an afternoon of games and contests, mostly on horseback: races, wrestling (where the objective is to knock your opponent of his horse), and the local specialty: a cousin of polo, played with the headless carcass of a goat. Horses cluster together as their riders lean and reach for the prize, and horses and riders fall over and get up again. The winner, sporting a magenta t-shirt, earned a roar from the crowd (sitting on the ground, on horseback, and on donkeys) and kissed the cheeks of a few of the girls in our group. Little boys, some probably only nine or so, sat on horses and donkeys. When we commented on how easily, how fluidly, they rode, our Kyrgyz guide said, "They are born on horseback." Most of the boys posed for cameras, grinning and showing off; one, with a dark green hat on, sat solemnly on his horse a little apart from the crowd. Tourists and locals and horses mingled, everyone smiling, grinning, shaking hands, cheering for the winners of each new game.
The yurts are made by hand, with wooden frames supporting woolen walls and felt mats on the floor. The opening at the top opens with a tug of rope to let in the sunshine, and closes again at night to keep us warm. So warm, despite the fact that the temperature dropped nearly to freezing. It rained the second morning, but just up the valley, fresh snow dotted the hills and mountains. If this is July, no wonder they don't spend the winters up there. Each spring, people drive their livestock – on foot – from the lower villages to their summer territory, bringing their yurts to set up and paying the government a fee per animal for grazing rights. It's a beautiful, harsh place to live, where water is pumped from the river and no plant larger than grass will grow.


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In 2009 I 'emigrated' to Spain. Unfortunately it didn't work out and I was back in Blighty pretty sharpish.
However, whilst there, I met a lovely lady called Josephine and after I returned to the UK we kept in touch. After the death of her husband, Josephine wanted to make their bedroom more girlie so to speak and asked me to do a painting for her.
When it was done I had been diagnosed with cancer, had an operated and begun chemotherapy. So I decided to hand deliver my painting to her.
Thankfully she loved it. I know because I didn't tell her I'd brought it with me and left it on my bed for her to find after I'd gone. Receiving her call in the taxi to the the airport was priceless.


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I totally need one of these with a rotating library of Keanu film trailers as the crown jewel of my T-shirt collection, which has grown considerably since this entry.
Right...that's all you need...MORE guys staring at your chest. ;)Posted by: Mikey2 on May 18, 2004 11:29 AM
LOL@mikey!!! who said it all, really... *grin*Posted by: bakednudel on May 18, 2004 11:36 AM
...naw, Mikey:
Krix wants the world to know that K is near and dear to her heart,
that's why she has to wear him on her chest...
Mikey 2 is so evolved!!! Impressive and lovable.
But maybe this is the way to convert more guys to Keanu-fandom. Those I-Robot guys are having insidious ideas over there it seems. ....but is it comfortable?Posted by: Tawny on May 18, 2004 01:12 PM
Hey, for some of us, shall we say, lesser-endowed gals, this is probably the only way we'd get any guys to stare at our chests.Posted by: Melissa on May 18, 2004 01:40 PM
That's another way for guys to stare, but then again you'll have the women too.. with Keanu there.. LOL! It would be way cool to wear one, with Keanu on it..
I wanna show Keanu not only is he near and dear ot my heart, but where he can rest his head, for the night, when he needs to LOL :)
hugs and God bless your friend
ok i realize yer TSHIRT COLLECTION thread, was waaaaay back in 2002..but ya mentioned $100 for a HAMLET TEE on ebay....they dont seem to be as much any more..but if someone didnt get a chance to get one back then..there seems to still be some listed today
>> http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&category=15687&item=3912493300&rd=1&ssPageName=WDVW
>> http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&category=778&item=3815482544&rd=1&ssPageName=WDVW
just sharingPosted by: Dawni on May 18, 2004 02:33 PM
Very cool. Me Want...Posted by: Keanuette on May 18, 2004 03:34 PM
keanu in motion, great, just... what about sound? that palpitant voice! and bateries? it remined me the matrix - human being change to this... cool! ... it is the life matrix! (Wait, Neo didn´t accepted it!)Posted by: vika on May 18, 2004 03:59 PM
LOL! LOL! @Mikey!...there are definitely SOME women who will not be able to effectively "display" it...nor will it change those women's present lives in ANY way...of course these women already struggle everyday as they try to eat in front of a "shelf."
...as for Neo...there is a "spoon" in the "life" Matrix...accept it!Posted by: shelz on May 18, 2004 05:43 PM
Bah, who needs boobies? I dig eyes!Posted by: Mikey2 on May 18, 2004 08:05 PM
5 billion years of evolution have brought us Mike! :-D
(I'm sorry, I'm STILL snickering at the 'Mikey2 is so evolved' part), LOL!
I think Krix doesn't need a t-shirt to attract attention :-) Altough I WOULD stare if she had Keanu attached to her chest ;-DPosted by: Julie! on May 18, 2004 08:40 PM
I would kill for A "constantine or MAtrix" T-shirt like that.Posted by: Keana on May 19, 2004 03:46 AM
Yeah, Julie!, then I went over to the Miketrix and re-considered that statement. But he is still lovable. He IS evolved, just not scarily over- evolved. :-DPosted by: Tawny on May 19, 2004 05:56 AM
I think Mikey is a "regular" guy Tawny..
...I *heart* Mikey :-)Posted by: shelz on May 19, 2004 07:01 AM
Too cool!! I think I'd need to have 2 of them. One to wear and show off and the other I could just have hanging around so I could stare at it myself! ;) I'm just weird like that sometimes! LOLPosted by: Lisa on May 19, 2004 09:54 AM
Aw... Tawny, you don't need to reconsider, LOL!
That was a great statement, and I couldn't agree more :-) It just made me giggle... A lot! :-D
Oh! Krix! I'm going to be bouncing with curiosity at that mystery parcel for days, LOL!
I need some tranquilizers or something ;-)


text:
The Promise Sleeping Pyramid
This is a portable Sleeping Pyramid System is designed to go over your bed to help your etheric body be protected while sleeping. When we sleep our bodies actually recharge like a battery off of the earth's magnetism.The larger Pytramid acts as a Faraday Cage and aids the body in recharging the aura. This device is also very functional as the ultimate Feung Shuei enhancer. It can even be used as a functional art piece. It can be set up in a well-trafficked area of your home to add a sense of calm. It can be used in a meditation room while inspiring a healing vibration.
This device is easy to assemble and it beautifully made and comes in a durable white powder coat paint. it comes in several sizes the most popular is 8ft base about 5 1/2 feet height or the 9 ft base that is about 6 feet in height. It comes with also comes with a stunning beautiful Gold Plated Capstone!
Please email RobThePromiseRevealed.com or Call me at +1- 530-925-3502 for personal assistance with this purchase.
Thank You
Learn More About Pyramids By clicking Here
The Promise revealed is one of the only worldwide distributors of pyramids, Orgone and Tachyon technology. The number one seller for several decades has been the amazing headgear pyramid called the Promise Gold. We have found that the Gold plated pyramid has a very strong detoxifying effect on the body as well as creating a much stronger output of the negative ion effect.
Head Gear Pyramids are 9″ titanium or steel based, plated with precious metals such as gold silver and copper. Everyone who has seriously endeavored to wear the pyramid headgear for any length of time has had remarkable results in the reduction of fatigue and stress. These pyramids have also been proven to help the body reach the alpha or meditative state more easily. How does the Pyramid Headgear work?
A large volume of magnetic concentration exists within the pyramidal boundaries which helps protect the human body from adverse effects of negative energies and radio frequencies because the Pyramid acts as a Faraday cage. With regular use, the pyramids help create a beneficial environment for your body to function properly because the Pyramid actually sings at a Fibonacci frequency. This creates a resonant field which harmonizes the body's magnetic field or Aura. This causes the endocrine glands to relax and to release the complex neuro-hormonal structures which the subjective consciousness registers as healing and bliss.
Learn More About Pyramids By clicking Here
Additional information
|9 × 9 × 9 in


text:
Grilled Italian Sausage and Peppers served in a bun and topped with marinara sauce, melted mozzarella cheese, and fresh basil. The perfect easy summer dinner! Summer Grilling We grill out almost every night during the hot summer months. One of
When fresh corn and tomatoes are at their summer best, this easy salad with creamy avocados that's simply dressed with a hint of Mexican jalapeño, lime juice, and garlic, is a healthy recipe that's sure to become the star side


text:
Manufacturer of Stainless Steel Wire Mesh
Our company is the major manufacture of stainless steel wire mesh,especially know as its best quality and excellent repulation.
Material: SUS302,304,304L,316,316L
Weaving: plain weave,twill weave,ductch weave.
Use : extensively used in mining,chemial industury,food industury and pharamaceutial industury.
Wire diameter: 0.025mm-2.0mm
Width: 0.5m-8m
Length of standard: 30m
Any size avalible as the requiements of the customer.


text:
Subintimal Crush of an Occluded Stent to Recanalize a Chronic Total Occlusion Due to In-Stent RestenosisInsights From a Multimodality Imaging Approach
Author + information
- Received January 23, 2017
- Accepted February 9, 2017
- Published online April 17, 2017.
Article Versions
Author Information
- Giuliana Capretti, MDa,
- Satoru Mitomo, MDa,
- Manuela Giglio, MDb,
- Mauro Carlino, MDa,
- Antonio Colombo, MDa and
- Lorenzo Azzalini, MD, PhD, MSca,∗ (azzalini.lorenzo{at}hsr.it)
- aDivision of Interventional Cardiology, Cardio-Thoracic-Vascular Department, San Raffaele Scientific Institute, Milan, Italy
- bCardiovascular Disease Prevention Center (CPC), San Raffaele Scientific Institute, Milan, Italy
- ↵∗Address for correspondence:
Dr. Lorenzo Azzalini, Division of Interventional Cardiology, Cardio-Thoracic-Vascular Department, San Raffaele Scientific Institute, Via Olgettina 60, 20132 Milan, Italy.
- chronic total occlusion
- in-stent restenosis
- intravascular ultrasound
- multidetector computed tomography
- optical coherence tomography
- subintimal
A 46-year-old man underwent percutaneous coronary intervention with a drug-eluting stent on the distal right coronary artery. Six years later, he was diagnosed with in-stent chronic total occlusion of the right coronary artery causing inferior-wall ischemia, and was scheduled for percutaneous coronary intervention.
The occlusion was long, with an ambiguous proximal cap, and interventional epicardial collateral channels from the circumflex (Figure 1A). An antegrade true-to-true attempt failed to cross the lesion, therefore a retrograde approach was undertaken. A Sion guidewire (Asahi Intecc, Nagoya, Japan) over a Corsair microcatheter (Asahi Intecc) was successfully advanced across the lesion and into the antegrade guiding catheter (Figure 1B). It was later noticed that the wire crossed the occlusion completely outside stent, through the subintimal space (Figure 1C). The decision was taken to crush the occluded stent and to implant a new drug-eluting stent in the subintimal space (Figure 1D). Three additional overlapping drug-eluting stents were subsequently deployed with good angiographic result (Figure 1E).
Two months later, coronary angiography (Figures 2A and 2B), multidetector computed tomography (Figures 2C and 2D), intravascular ultrasound, and optical coherence tomography (Figure 2E) were performed, showing persistent optimal result in the right coronary artery and particularly at the site of subintimal crush of the occluded stent. The patient remained asymptomatic throughout follow-up.
Although in-stent chronic total occlusion is usually recanalized using a true-to-true lumen approach (1), subintimal crush of the occluded stent represents an alternative strategy. Invasive imaging in this setting is useful to confirm optimal crush of the occluded stent, and good expansion and apposition of the newly implanted stent in the subintimal space (2).
All authors have reported that they have no relationships relevant to the contents of this paper to disclose.
- Received January 23, 2017.
- Accepted February 9, 2017.
- 2017 American College of Cardiology Foundation
- ↵
- Azzalini L.,
- Dautov R.,
- Ojeda S.,
- et al.
- ↵
- Roy J.,
- Lucking A.,
- Strange J.,
- Spratt J.C.


text:
Balance your hormones, lose the weightGet Started
I want...
January 23, 2012 By Leanne Vogel July 1, 2015The potential coconut sensitivity that I mentioned on Friday = false alarm. Thank goodness. I spent most of the weekend panicking that my love for coconut had eventually led to a sensitivity. (When you eat too much of one thing, especially when you're prone to allergies, your body can begin to develop a sensitivity to it) What I neglected to think of was that having something high in fat after being on a cleanse for a week; regardless of what type of fat, would lead to stomach pains. Sure enough, I had coconut on Sunday and didn't have any type of reaction. Boooya! But there's a lesson in all of this. Maybe I should try to limit coconut to a couple servings a week just so I don't develop a sensitivity to it. …. Just after I finish these delectable bundles of ginger spice goodness, kay? I'm starting up with spin classes again this week and knew that the added cardio would require me to eat more. Instead of grabbing for a LARABAR, handful of store-bought granola, or a packet of trail mix, I wanted to create with something I could really feel good about eating. Something that was,
This entry was tagged: coconut flour, ginger, grain-free muffin, protein snack
Nutrition educator + keto enthusiast. I want to live in a world where every woman loves her body, nourishing fats are enjoyed at every meal, and the word "restriction" isn't in the dictionary.


text:
Blowing in the wind
From the Senate in the 1970s to the presidential campaign trail in 2020, Joe Biden has a long record of going where political pressures push him—and right now they're pushing him aggressively leftward
From the Senate in the 1970s to the presidential campaign trail in 2020, Joe Biden has a long record of going where political pressures push him—and right now they're pushing him aggressively leftward
Massive blast kills dozens, injures thousands, and levels neighborhoods
The U.S. has halted asylum hearings, and migrants hoping for entry are in limbo
The pandemic divides Americans—even Christians
Andrée Seu Peterson / Janie B. Cheaney / Joel Belz / Marvin Olasky / Mindy Belz / The Editors /
Education / Technology / Lifestyle / Sports / Health / Medicine / Law / Business / Science / Religion / Money / Media
Dean's List / Metro Minute / Snapshots of China / Sophia's World / Whirled Views /
Television / Movies / Books / Q&A / Music
Quotables / Human Race / Quick Takes / News
I recently wrote a story about homeless women. The main purpose was to highlight one of the most vulnerable populations in our country, particularly at a time when all sorts of groups are calling attention to gender inequality and social justice issues.
But I also want to emphasize one more point: The women in my article have suffered great injustice and abuse and deserve help and compassion, but they're also not helpless victims. They're neither weak nor lazy. They admit they've made some very bad choices, but they cling to life, hope for goodness, and long to help others. They're beautiful women, and they're survivors.
Take 64-year-old Mary, for example. I first met Mary last year while volunteering with a nonprofit that feeds the homeless in Venice Beach. The homeless in Venice are a mixed bunch, but a good chunk of them are young, drug-addicted vagrants and artists who for now seem content with their lifestyle. Mary, though, was a single woman with a walker, a stooped back, no upper teeth, and a creased face. She had osteoporosis and kneecaps that sometimes popped out when she landed on her feet too hard. She didn't seem to have a drug or alcohol problem, and her mind was clear and sharp. She clearly didn't belong there, but had somehow, by sheer force of personality, wiggled her way into the Venice homeless community and gained a title as "Grandma." The homeless are fellow human beings with a heart: The young men took care of "Grandma," made sure she had food to eat, and paid her a few bucks to sell trinkets at a booth on the beach walk.
When she gets her monthly disability check, Mary blows most of it by taking her buddies—Bear, T, Doose, Johnny, and the gang—out to a sit-down dinner at a restaurant. She spends the rest of the money helping them buy little things they need, such as new batteries and extra hoodies. Her money runs out in four days, but Mary says she doesn't regret it, because "these kids deserve to have at least one meal where they can sit down and be waited on without worrying about homelessness." Spoken like a true grandma.
Be warned if you take her out to eat, though. This 137-pound lady can in one sitting down 10 giant, cheese-laden, sour cream–drenched tacos on all-you-can-eat $5 Taco Tuesday nights. The two times my boyfriend David and I took her out to dinner, she ordered a 14-ounce bone-in ribeye with clam chowder and thick-cut steak fries. As we watched with wide eyes, she polished off the massive meat slab, then licked the marrow off the bone. It was one expensive meal (David and I had cheesecake and water), but the delighted expression on her face was worth it. For Mary, this was a rare treat, and she was determined to enjoy every sip and chew, even if she had to skip a couple of meals after that.
Later, when Greg (the photographer for my story) offered to treat her to "anything on the menu," I forgot to warn him. Greg later sent me a picture of Mary holding up her $29 steak next to her $7.50 margarita (extra salt). "Mary is in her happy place," he texted me, adding an "astonished face" emoji. He himself had a hamburger and a glass of water.
Every homeless person I've met has an interesting story to tell, and Mary's story is tragic: She says her mother left the family when Mary was 9 months old. When she was 6, she almost died from rheumatic fever. She was raped when she was 10. Ten years later, her father died as a crewman on the SS Edmund Fitzgerald on Nov. 10, 1975, and she still carries his picture around and can rattle off every obscure detail about that ship. She hitchhiked to Southern California when she was 17, worked as a roadie for a rock 'n' roll band for three years, then got pregnant with her then-boyfriend. That man beat her up for getting pregnant, so Mary fled to Pennsylvania to deliver her oldest son. There, she met and married Allen, the "love of my life," and moved with him to Wisconsin.
Mary gave birth to two more children with Allen—one daughter, one son. While she was nine months pregnant with her third child, the doctors diagnosed Allen with leukemia, and he died within a month. Mary sunk into a deep depression and spiraled into drugs. Eventually, realizing she was in no condition to take care of three kids including a baby, she gave her kids up for adoption. "I regret that decision every day of my life," she told me, eyes glistening. "I gave my babies away! Just because I couldn't handle it. I thought I did the best thing I could for my kids."
Over the next four years, Mary found her way back to California, an 87-pound, homeless addict. Then someone on the streets offered her help, and she took it. She went through detox and says she hasn't used drugs ever since. Desperate for work, she lied her way into a technician job with Warner Bros., until the night in 1994 when she says a gang of men raped her and tossed her half-dead into an alley. Two homeless men saw her and saved her life.
As she told me her story, I teared up too, and Mary gently brushed the drops off my face with her fingertips. There is much to Mary's story beyond homelessness: abandonment, sexual abuse, domestic abuse, loss and grief, love and motherhood, injustice and mercy. I was amazed that one woman could bear so much in her 64 years on earth and not be comatose with despair and bitterness.
No, Mary is determined to wring the most out of life, even as a homeless woman. I admire her resilient spirit, which is a gift from God. This New Year's Eve, she took the two-hour metro ride to Pasadena with her sleeping bag, a big orange jacket, a thermos full of hot chocolate, a beanie, and a folding chair to camp out in wait of the 2019 Rose Parade. While I watched the parade on TV beside a fireplace the next morning, she was out among the crowd in 30-degree weather, soaking in the live entertainment. She texted me later saying she felt like a sardine squeezed into a can, but she had great fun.
People sometimes paint the homeless as having a sense of entitlement and demanding free housing and services, but that's mostly untrue regarding the homeless people I've met. A few nights before Christmas, David and I went to visit Mary at her new shelter in Santa Monica. Some people who knew about Mary had been creating a fuss to the city about a 64-year-old woman out on the streets, and now three nonprofit teams are trying to help her find housing. Mary was thrilled—not just about the possibility of housing, but also the idea that random people have been advocating on her behalf. "You know what this means?" she told us, fists clenched and trembling in excitement. "That means ... I'm endorsed!"
But as she told us of her upcoming birthday and Christmas plans, I felt sad, realizing that while she was making plans to make the occasion special, they all involved being with a group of strangers, such as a free dinner at a comedy show club. It made me realize a common trait homeless people share: They're alone.
Before we said goodbye, David and I handed Mary a Christmas gift. I had baked her banana bread with chocolate chips and cranberries, and David and I had picked out a velvety, burgundy sweater for her. As soon as Mary unwrapped her gift, she clasped the fuzzy sweater to her chest. "Ooooooh! I love it, I love it!" she cried. Then she threw her arms around me in a tight hug, and did the same for David before scurrying back to the shelter: "I have to go before I ... before I …" she said, sniffling.
That Mary. Stubborn as always, a tough cookie. She can't even let us see her cry.


text:
Hi everyone,
I have just started a loft conversion in Bromley with a L shaped dorma creating a master bedroom en-suite i will keep you all up to date with regular blogs and photos. Here is some of what is completed to date: stripped the old roof, fitted ridge beam, floor beams and just starting the dorma. Tomorrow we will be fitting the roof beams and finishing the dorma.
I am waiting for finished photos of the refurbishment i have completed in station rd, Meopham should be getting them in the next few days. I have also done a mansard roof conversion in Wimbledon since my last blog.
Daniel Bishop


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Posted :
30 September 2013 12:25:35
Rank: Beginner Level 2
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Joined: 30/09/2013
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Location: Scotland
Morning All
A couple of books that you might find useful
I have found both of these enormously helpful (and just plain interesting) over the years.
I think they are both out of print now but there are plenty of copies available second hand. Do shop around though as I have seen the James Lees book offered for prices ranging from £35 to £400+.
Cheers, Frazer
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Posted :
24 July 2019 17:37:04
Rank: Semi-Pro Level 1
Groups: Registered
Joined: 17/04/2010
Posts: 52
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Location: Bradford
Came across this today:
Sovereign of the Seas, 1637: A Reconstruction of the Most Powerful Warship of its Day
Hardcover – 28 Feb 2020
by John McKay (Author)
Not out till 2020, but looks like it could be very useful.
In the meantime I have ordered:
Sovereigns of the Sea: The Quest to Build the Perfect Renaissance Battleship
by Angus Konstam
For a bit of light reading.
Building: R2D2 and Sovereign of the Seas
Subscribed: Bounty
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July 24, 2019
The 28th National Vocational Education and Training Research Conference 'No Frills' took place in Sydney from 10-12 July and was co-hosted by TAFE SA. IBSA attended the annual conference as it was an opportunity for researchers, practitioners, providers, government and industry representatives to come together and share knowledge, ideas, insights and solutions surrounding Australia's Vocational Education and Training (VET) sector.
Minister Cash opened the conference and shared how skills and vocational training are central to her portfolio as they lead to employment outcomes as well as skills for life. The key note presenter, Dr Fiona Kerr, a prominent thought leader and Neural and Systems Complexity Specialist at the University of Adelaide delivered the conference keynote address. Her address discussed the neurophysiological effects of interaction between humans and technology and how it relates to learning and the adaption of new technologies.
IBSA CEO Sharon Robertson also emphasised the importance of skills development in meeting emerging industry needs and enabling future generations to build capabilities throughout their career in her presentation 'Shaping the skills of our future workforce'. Click here to view her presentation.
During the three-day long conference IBSA met key stakeholders and gained additional insights into the student journey, and the important role VET plays in assisting workers to become lifelong learners, and continue to grow and evolve with their jobs in the changing world of work.
IBSA also attended the Conference Dinner which was held at the Graduates Restaurant at TAFE SA Regency Park Campus. It was great to see students involved as part of the program from food preparation, service – even the table centerpieces were created from chocolate as final assessment pieces. It was also interesting to hear how Coopers Brewery maintains its competitive advantage while staying true to its family heritage and core values.
Key Takeaways
Some of the key takeaways from the Conference were on how VET supports lifelong learners through:
- Developing learners' digital skills to productively use new and emerging technologies in response to the challenges of an increasingly complex and changing world,
- Understanding how the VET system will need to transform itself from initial skilling for jobs to formulate training to open job prospects for existing workers through upskilling and reskilling, and
- Supporting learners to develop a 'lifelong learning mindset' through the development of soft skills including adaptability, perseverance, resilience, curiosity, responsiveness, experimentation and initiative.