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Everyone cannot afford an expensive PR firm to help them promote their video, but you can have the next best thing. Just do what PR would do yourself. First, look at everything from your audience's vantage point, then get to work to make sure your audience sees it – while making it all look even better for them.
Finally, as you become more popular, send out press releases about the things you're doing outside of YouTube. Get active within your community, and become a well-known name brand throughout the internet and not just where your videos reside.
Any time you put up a video you need to get feedback from your viewers so that you can improve it each time. And the best way to get feedback is just to ask. It's amazing, but when you take the extra step to ask for feedback, you'll be more likely to get the feedback you want. You can even be specific to the type of feedback you want. For example, if you got the new sound equipment you can ask, "Let me know if you like how this video sounds."
The more specific you are when asking for feedback from viewers, the better and more useful the feedback will be. Here are some tips.
If you want feedback you're going to have to ask for it, every single time you post a video. People will only comment on what they want to if you don't ask for specifics. If you are good about giving feedback with other video creators, you'll establish a good relationship with them, and they'll do the same for you.

subset: commoncrawl


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Online design banner jobs
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...site de forma automática na data e hora programados pelo criador do grupo com contagem regresiva. A remuneração do site será: 1. pagamento de mensalidade 2. anúncios no banner com redirecionamento 3. posicionamento de destaque do grupo criado 4. Criação de grupo privado -----------------------------------------------------------...
...alguns produtos, precisaremos de apoio pontual em content marketing, nomeadamente design para diversos meios online, em diversos formatos tais como: Imagens para Artigos de blog (Header Image e footer banner image) Imagens para Email Mkt ((Header Image e footer banner image) Templates de Ebooks (valor por hora ou por página ou por palavras) Estes
...Criar dentro do Template Responsivo que já temos alguns modelos que compramos no site: [login to view URL], uma área de Login e senha como modelagem na vesão Desktop / Online. • Segunda Fase = Começarmos a Versão Beta do Aplicativo. Obs.: Sou Designer e posso ajudar muito no Projeto. Já tenho o Logotipo e criei alguns Ícones. E...
Quero construir uma loja online para venda de produtos tecnológicos (informática, comunicações, multimédia, iluminação a leds, segurança electrónica). necessito de um design uniforme (logo, cores, banner loja, banner facebook)
Preciso do Design e plataforma para administração. Será apenas 01 site com dois segmentos: 1º loja virtual normal / 2º site para locação online, ou seja, a pessoa irá escolher o produto, efetuará o pagamento referente ao aluguel + frete pelo site (cartão). Preciso de uma plataforma única para administrar esses d...
Criar site de cursos online, onde o aluno fará inscrição no curso, terá sessão de provas, e opção de emissão de Exemplo do site: [login to view URL] e [login to view URL] . Opção de entrar com login do facebook, emissão automática de email ao fazer o cadastro, opção de pagamento, op&cced...
...noite Ricardo, Preciso de um website simples em Wordpress, nas línguas PT e EN, com 3 Páginas: 'Sobre nós', 'Contactos' e 'Siga-nos'. O site será posteriormente uma loja online, mas neste momento preciso de algo para apresentar a empresa a potenciais fornecedores, daí a importância de o ter em 2 líng...
...Apresentar a empresa, catálogo de produtos, comercializar produtos online, receber contatos. Referência [login to view URL] Informações adicionais - Design Responsivo em HTML5 Briefing para desenvolvimento PáginasPosição Seção do site Módulo Descrição do conteúdo 1 Banner de Destaque...
...Education Category - For Students and Teachers - Online Testing Platform and Diagnostics - Need a Logo and Several Image/Text Frames for Concepts that will scroll like a storyboard in the banner of home page. Very Similar to Amazon AWS banners, New Concepts and Features will be displayed in the banner Tips - Keep it simple but still appealing. Don't
i need cover pic banner design for my online meat delivery business
Requirements: E-Commerce website design Business name: MeBlinds Logo: Attached Style: Simplicity and clean What we do: Custom DIY Window covering/blinds for the Australian market. The emphasis is on "me" where it's blinds for Me or custom made for me. Target Audience: Mainly homeowners that prefer to save money when changing their blinds
-s...Facebook and Instagram ad banner creation & More - Online Reputation Management - Group posting activity - Online campaigns suggestion - Post in Online directory Like Justdial /Yellow page & More available sites - Advertisement Banner in other popular site - Email Mail Marketing - Offer Banner Note: Design and Advertisement Banner...
...INCLUDE I need a new website. I need you to design and build my online store. for my TEA PRODUCT . AND I NEED FILLOWING THINGS TO BE DONE. Web Designing and Development Proposal Page Templates · Design Templates ( · User Visibility research and Layout Engineering Template Layout Banner · Size according to spe...
I am needing a logo and banner created ASAP for online jewelry. Looking for a boho design using a mandala and earthy tones for colors. I will need a square logo and 1200 x 600 banner.
I need a logo designed. Need logo for website... Something like banner plus small logo for profile picture Its online Aroma shop, which will include essential oil, diffusers, smoke cones, smoke sticks and other aromatic products. Design need whole words "home Essence" Logo should represent the nature of business.. Profile picture logo can be "HE"
I'm starting an outfitting business which consists of selling trips, also called packages. I need someone to design my website which will be composed of only 4 or 5 pages. The key features of the website will be; - Online purchase (i.e, cart management, check out, Paypal payment, etc.) - Discount code - There is a fix number of the same package. When
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...Job Postings** Web design/ graphic design/ artistry/anime designer ( Temp/Part-time) I have two separate jobs i'm looking to have filled for two separate companies. Message me privately here or send resume and a blurb as to why you would be a good fit with copies of work in email. ( lauraelizabethkerr@[login to view URL]) Job 1: An online company is looking
...follows: Front end a. Website design, development and functionality coding along with admin control panel to manage the portal contents. b. Requirement Analysis and Study c. Database Design d. Third Party xml feed integration for Christian search e. Integration of Google AdSence ad feed. f. User Management g. Banner / classified management h. Website
Por favor, Cadastre-se ou Faça Login para ver os detalhes.
We need a banner created for a website with a 3 word logo. No sign necessary. It needs to be downloaded in the correct format which can be easily transferred to a website powered by [login to view URL] 3 words need to have a dystopian feel, with bold colours and strong lines. The background banner needs to be space/galaxy themed. This should be completely
Banner Design and Product Design.. improve Logo ( minimum Five Variant Design for Vertical and horizontal sizes with new innovation building material online market place)
We are an online medical specialist directory - [login to view URL] We would like to introduce completely new layout that would better show the existing content as well as new content for specialist profiles - [login to view URL] * Make the page shorter, better use the space (more columns etc) * The
...square-metre, allowing customers to calculate bespoke Banner sizes - The counter on product ordering page to make the customer aware of the deadline for ordering and submitting their artwork. (Check SoloPress Website) - Print Product Catalogue - Matrix, Lookup & Large Format Pricing - Online Design Tool (if customer don't have there own artwork) with
I need a new website. I need you to design and build my online store. I need a do-it-yourself Banner design website. Customer could lay out a simple banner and send it in as an order or request design help and we would do the layouts. Website needs to be secure checkout.
We have an online course site which build by a wordpress template: [login to view URL] We want to do some re-design to improve the wordpress site and We need an expert to make the wordprese be faster. The Design details is at: [login to view URL] Other Requirements:
Need a logo and web banner for a newly launched business that deals with small business consulting and business coaching. The name of the company is Steps2Success. The colour of the web site will be predominantly green (lime green). The online examples that catch my attention are at: [login to view URL] and the 'E' in the purple steps at https://bit
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[login to view URL] is the Best Online Casino Reviews website - We need a full width header banner and an animated logo - Header image(PNG) 2000px width x 400px height - Animated logo(GIF maybe?) 150px x 150px The header image should be something repetitive related to online casino, gambling, but not something with cards and dices only, something
We are a new company selling UPS Uninterruptible Power Supplie...selling UPS Uninterruptible Power Supplies for servers and computers in an online store and looking for a new logo. The company name is Critical Power Solutions Would like a long banner type logo and also a more square one so we can use on stationary etc Both with the same design.
Good Day I am setting up and online shop and I need a talented Marketer and Designer that will create a banner that best fits my marketing schemes and design and feel of the website. In total I will be needing 6 banners all of which will be placed in the website. If you think have the skills to do this, please apply.
I need a landing page (home page only) design in PSD. Details: > It's an E-commerce website and domain is [login to view URL] > Title: Importers & Exporters, Manufacturers, Suppliers from the world's online B2B Marketplace - Stm Corporation > Description: The main purpose of this website is >> Getting sellers >> Sell products/ wholesale and re...
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Dear Friends , Our company "Import In EU Trade Directory" would like a Professional Linkedin Cover Banner . Our site is an online Business directory .Where companies submit their businesses for promotion porpose especially in EU region. Our Prime Objective : We want to encourage linkedin visitors to visit our website and submit their business.
Dear Friends , Our company "Import In EU Trade Directory" would like a Professional Linkedin Cover Banner . Our site is an online Business directory .Where companies submit their businesses for promotion porpose especially in EU region. Our Prime Objective : We want to encourage linkedin visitors to visit our website and submit their business.
I'm doing online business, selling kind of products such as sports products & rubber stamps. Looking for a designer to design some product photos, banner, cover for website and social media such as FB/instagram
...that can create custom banner designs with little supervision. This is going to be a long term project. Hi We are a software development team and recently we just built a banner design software and we need graphics designers to help us build out templates using the software. It is quite easy to use just like Canva and other online graphics tools. This
Design a logo for a new online Store "Grets" Grets Technology sneak peak of site is here [login to view URL] ideas modern fresh. crisp nothing like word art will win. good as black and white and clour Australian owned and operated. confident we need all files after this is closed font name and font file or link to buy it hex
...looking to do is design a logo / symbol for my YouTube and online brand. My username on everything is Ebontis, but also a graphical representation that fits everything's circle shaped brand symbol. I also would like to incorporate a cheetah as my symbol in some way if possible, but I'd prefer something unique as I've seen some options online but prefer
I would like someone to create attractive banners design using the clients images to use for promotions on website. please make sure that the banners flow with Website theme color and design standards (attached website screenshot) Size of banner 1900*600px #### WHAT TO INCLUDE #### 1- Select any of the client's images attached. 2- Use the following
Web Design PHP/CSS/BOOTSTRAP Project Requirements Required skills: HTML, PHP, CSS, SQL, BOOTSTRAP I have all the functionality coded in PHP and this just needs to be plugged in to a basic dashboard so it displays well and looks professional. I also have an admin panel which is quite minimal and needs some CSS work. Details: • Incorporate all php functionality
...mascots and website banner. Feel free to take on the concept! Minimum requirements is a 2D illustration; added bonuses would be advertising skills, converting to computer image. Style wanted: Chibi, Anime, Cartoon Artistic job: 2 unique mascots (fish, crab, lobster, clam), aesthetic design Job: create 2 banner ad designs, logo design and website header
Hi Zuomin X., I have a small update to the header banner on our website. I hope this will be a fast and easy change for you. The banner currently says "BUY NOW during online pre-sale / orders expected shipping from 15th June." We want to make 3 changes to the design: (1) change the font our Gordita font (2) increase the font size (increase height
I need a new website. I need you to design and build my online store. New ECOMMERCE website with Multi vendor,classified both including, Secure Web Design, Free Email A/C'S, Unlimited pages, Logo, SEO submission, Free Domain, Banner ,Registration form,Contact us form,unlimited Image gallery, CMS /Dynamic, Optimised Coding with 3 samples, W3C
...baru Membuat dan mendesainnya Toko online [login to view URL] Job ads list Designer Grafis untuk Website Pro Solusi Webmedia Jakarta Utara Pro Solusi Webmedia Persyaratan Syarat: Punya minat dan memang tertarik dengan website. Menguasai Photoshop dengan baik. Mampu menggunakan Illustrator, Corel Draw, atau software design lainnya. Bersedia untuk belajar basic
...woocommerce integrated with rental. So people can book a inflatable model for a certain period. Can you integrate Woocommerce with rental in our website [login to view URL] (design of total website stays as it is. (so the productpages as [login to view URL] and [login to view URL] will change


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Introductory Quantum Mechanics 4th Edition Richard The graded exams are available for pick-up in my office any time this week. Buy Introductory Quantum Mechanics 4th Edition on ✓ FREE. but doesn't explain how to do half of them and doesn't have a solutions manual.
Introductory quantum mechanics liboff solution manual - Have a great winter holiday break---you earned it!!! Introductory quantum mechanics liboff solution manual. Jim Summers. Loading. Unsubscribe from Jim Summers? Cancel Unsubscribe.
Physics 139B Home Page - SCIPP For the distribution of ages in Section 1.3.1: (a) Compute (b) Determine for each j, and use Equation 1.11 to compute the standard deviation. Final exam solutions are posted to Section III of this website. The graded. Introductory Quantum Mechanics, 4th Edition by Richard L. Liboff.
Introductory quantum mechanics - -           Lectures: Tuesdays and Thursdays, -- pm, ISB 231 Introductory Quantum Mechanics, 4th Edition by Richard L. Griffiths - Introduction to quantum -. the Physical Sciences, A solution manual is available to instructors. solution manual mary l boas.
QM Homework Solutions (c) Use your results in (a) and (b) to check Equation 1.12. The following table contains the homework solutions for Quantum Mechanics 1. They are taken from Introductory Quantum Mechanics by Richard L. Liboff and.


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After 41 years of service as pastor of the United Methodist Church, including the last seven years at Kenwood Park United Methodist Church, Pastor Michael is retiring June 30.
He reflects: "As I look back over my years in ministry, it is the people I most remember. All of us came together in our shared journey through this miracle of life. I am thankful for having known them. It has been a privilege to minister in Christ's name with them all."
Cards can be mailed to:
Kenwood Park United Methodist Church
175 34th St. NE, Cedar Rapids, IA 52402
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This week I made spring rolls for some friends and family. I love the crunch you get from the veggies and the fresh herbs in these rolls make this so delicious! Serve with a peanut sauce, teriyaki, or a sweet chili sauce. This makes a great appetizer before the main course. I also love to just eat it as a bowl without the rice paper wrap and its so light and refreshing on a warm day.
Vegan Note: You can make these with tofu or just add extra veggies in place of the shrimp.
Yield: 10 rolls
7 ounces rice noodles
10 rice paper rounds
2 carrots
1 cucumber
1/4 head of cabbage
30 shrimp fully cooked frozen, medium size, peeled and deveined
fresh mint
fresh cilantro
sesame oil
Cook the rice noodles per directions, run under cold water, drain and set aside.
Wash all produce, peel the carrot and cucumber, cut each into thirds, and then slice into thin strips. Remove the core and the outside layer of the cabbage and discard. Slice very thin. Wash the herbs and pat dry.
Take the shrimp and run under cold water in a colander for 5-10 minutes until thawed. Remove the tails and set aside.
Take a large pot or bowl and fill with warm tap water. Take one rice paper round and soak in water 15 seconds, take out let excess water drip off and lay on a cutting board or plate. Take a small handful of the noodles, about 1/2 cup, and place in the middle. Layer on top just 2-3 pieces of the carrot and cucumber, and just a little cabbage. Add 3 shrimp and the fresh herbs, just 2-4 pieces of each cilantro and mint. Depending on size and preference for how much you like to add, I like to chop up the herbs and add sprinkle them on top. Then last just drizzle a small amount of the sesame oil all along the top.
Taking the sides of the rice paper and fold them in towards the middle. Then fold up the bottom to cover all the filling and roll the rest up until you have a completed spring roll. Repeat this process until all rolls are make and enjoy!
I have had trouble with delivering these because sometimes when you put them altogether in a container, the rolls can stick together. To avoid this I wrap each one individually with plastic wrap or use wax paper in between the layers and that helps.
Note: If you refrigerate this for more than an hour the rice paper tends to get hard and doesn't stay soft, so don't make them too far in advance.
Would you like to see more recipes like this one on the blog? What do you like to add into your spring rolls? Let me know in the comments below and happy cooking!
xoxo- Julie


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Greenville, TX Coach Factory
YP - The Real Yellow PagesSM - helps you find the right local businesses to meet your specific needs. Search results are sorted by a combination of factors to give you a set of choices in response to your search criteria. These factors are similar to those you might use to determine which business to select from a local Yellow Pages directory, including proximity to where you are searching, expertise in the specific services or products you need, and comprehensive business information to help evaluate a business's suitability for you. "Preferred" listings, or those with featured website buttons, indicate YP advertisers who directly provide information about their businesses to help consumers make more informed buying decisions. YP advertisers receive higher placement in the default ordering of search results and may appear in sponsored listings on the top, side, or bottom of the search results page.
Tandy Leather
Serving the Greenville area.
- (800) 433-3201
Morris Kaye & Sons
Serving the Greenville area.
- (888) 224-3005
Group USA and Camille LaVie Women's Apparel
Serving the Greenville area.
- (888) 293-9461
1. Coach
490 Bending Oaks TrlRockwall, TX 75087
2. Coach
I visit the Allen Outlet once a week on Friday's after work and I find that I get the first digs on all bargins. It's a wonderful place to walk around and sneak in and out of the many shops they offer. The salesfolks are extremely nice and the atmosphere is just heavenly! My favorite is Wilso…
3. Ribbon Outlet The Factory
421 Industrial Dr ESulphur Springs, TX 75482
4. Uniform Factory Outlet
820 W Stacy Rd Ste 112Allen, TX 75013
5. J. Crew Factory Store
820 W Stacy RdAllen, TX 75013
6. Uniform Factory Outlet
301 Tanger Dr Ste 220Terrell, TX 75160
7. J.Crew Factory
820 W Stacy Rd Ste 644Allen, TX 75013
From Business: Get classic J.Crew style from J.Crew Factory. Buy discount men's clothing, and women's clothing. Find great deals on sweaters, dresses, suits, shoes, accessories and jackets.
8. Furniture Factory Outlet
1405 Exchange DrRichardson, TX 75081
9. Master Hatters Factory Outlet Of Texas - CLOSED
2355 Forest LnGarland, TX 75042
10. Group USA and Camille LaVie Women's Apparel
305 dollars Hellobridals Vera Wang Faye
11. Heal USA
Beautiful shoes and amazing quality. I ordered a pair of these for my girlfriend and she loves them and I do to.
12. Ross Dress for Less
6834 Wesley StGreenville, TX 75402
13. Half Price Handbags
223 Main StSulphur Springs, TX 75482
14. Uptown Cheapskate
You have some beautiful things! Staff was very helpful, very good deals on name brands!
16. Elie Tahari
820 W Stacy Rd Ste 442Allen, TX 75013
17. Vera Bradley
858 Watters Creek BlvdAllen, TX 75013
18. Handbag Corner
430 Cedar Sage DrGarland, TX 75040
19. Claire's
2063 Town East MallMesquite, TX 75150
20. Loft
820 W Stacy Rd Ste 240Allen, TX 75013
21. Eddie Bauer
865 Watters Creek BlvdAllen, TX 75013
22. Fossil
820 W Stacy RdAllen, TX 75013
23. Avenue
811 E Yellowjacket LnRockwall, TX 75087
24. Jen Wall's Purses and Jewelry
500 Wentworth drRichardson, TX 75081
25. Snap 'n Zip LLC - CLOSED
3302 Ashwood LnSachse, TX 75048
From Business: Snap 'n Zip is an emerging online boutique that sells authentic contemporary designer handbags/purses and related fashion accessories at bargain prices. Our products are 100% authentic, on-trend and carefully selected from major retailers in high end malls or premium factory outlets (thus they have a very limited quantity)…
26. Skagen Denmark (formerly Skagen Designs Ltd.)
901 S Central ExpyRichardson, TX 75080
27. Wilsons Leather
2112 Town East MallMesquite, TX 75150
28. Nine West
820 W Stacy Rd Ste 184Allen, TX 75013
29. Torrid
301 Tanger Dr Ste 211aTerrell, TX 75160
From Business: Torrid's collection of trendy plus size clothing - including the latest apparel, plus size lingerie and accessories - is inspired by and designed to fit young, stylish women who wear sizes 10 to 30. From party-ready cocktail dresses and curvy maxi dresses to shirts-and-jeans outfits, our stores and online shop are the best…
30. Brandi Rose Country Store
5684 County Road 437 Lot 31Princeton, TX 75407


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Biotin is an essential vitamin that binds streptavidin or avidin with high affinity and specificity. As biotin is a small molecule that can be linked to proteins without affecting their biological activity, biotinylation is applied widely in biochemical assays. In our laboratory, IgM enzyme immuno assays (EIAs) of µ-capture format have been set up against many viruses, using as antigen biotinylated virus like particles (VLPs) detected by horseradish peroxidase-conjugated streptavidin. We recently encountered one serum sample reacting with the biotinylated VLP but not with the unbiotinylated one, suggesting in human sera the occurrence of biotin-reactive antibodies. In the present study, we search the general population (612 serum samples from adults and 678 from children) for IgM antibodies reactive with biotin and develop an indirect EIA for quantification of their levels and assessment of their seroprevalence. These IgM antibodies were present in 3% adults regardless of age, but were rarely found in children. The adverse effects of the biotin IgM on biotinylation-based immunoassays were assessed, including four inhouse and one commercial virus IgM EIAs, showing that biotin IgM do cause false positivities. The biotin can not bind IgM and streptavidin or avidin simultaneously, suggesting that these biotin-interactive compounds compete for the common binding site. In competitive inhibition assays, the affinities of biotin IgM antibodies ranged from 2.1×10−3 to 1.7×10−4 mol/L. This is the first report on biotin antibodies found in humans, providing new information on biotinylation-based immunoassays as well as new insights into the biomedical effects of vitamins.
Citation: Chen T, Hedman L, Mattila PS, Jartti L, Jartti T, Ruuskanen O, et al. (2012) Biotin IgM Antibodies in Human Blood: A Previously Unknown Factor Eliciting False Results in Biotinylation-Based Immunoassays. PLoS ONE 7(8): e42376. https://doi.org/10.1371/journal.pone.0042376
Editor: Sylvie Bisser, INSERM U1094, University of Limoges School of Medicine, France
Received: March 29, 2012; Accepted: July 4, 2012; Published: August 3, 2012
Copyright: © Chen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This study was supported by the Helsinki University Central Hospital Research and Education Fund, the Sigrid Jusélius Foundation, the Medical Society of Finland, the Academy of Finland (project 1122539), and the Orion-Farmos Research Foundation Grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Biotin, also known as vitamin B7 and vitamin H, is a water-soluble micronutrient required by most organisms. In mammals, biotin acts as a coenzyme for four carboxylases: propionyl-coenzyme A (CoA) carboxylase, pyruvate carboxylase, methylcrotonoyl-CoA carboxylase, and acetyl-CoA carboxylase. The first three are located in mitochondria and the fourth in the cytoplasm. These enzymes catalyze critical reactions in the intermediary metabolism of gluconeogenesis, fatty acid synthesis, and amino acid catabolism [1], [2]. Therefore, insufficient biotin intake may lead to a number of clinical abnormalities, including hair loss, dermal rash, growth retardation, neurological disorders and a higher vulnerability to infections [1], [3]–[5].
Biotin deficiency is rare among constitutionally healthy people as this nutrient is widely distributed in foods and also amply synthesized by the intestinal flora, meeting with our low daily requirement (approximately15–70 µg/day) [1], [2]. However, biotin deficiency can ensue upon excessive consumption of raw eggs, due to its constituent, avidin, binding biotin at high affinity in the alimentary tract, and preventing its intestinal absorption [3], [6]–[8]. In the human body, only free biotin can function in metabolism, whereas most of the dietary biotin is protein-bound [2]. Biotinidase, a hepatic enzyme synthesized in the liver and secreted into the blood, is responsible for processing protein-bound biotin and recycling biotin [5], [9]. In human serum, biotin circulates in free form or protein-bound, for uptake by cells and tissues including liver cells, cerebral capillaries, basolateral membrane vesicles of placenta, and peripheral blood mononuclear cells [1], [2].
(A) Layout of biotin IgM indirect EIA (Bio-EIA). (B) The distribution of EIA absorbance values of 612 adults' serum samples of Group 1 and Group 2. Five-pointed star: HRP conjugates.
Due to its small size, biotin can be covalently linked to a number of proteins without affecting their biological activity. As biotin binds avidin or streptavidin with extremely high affinity and specificity, biotinylation of proteins and macromolecules is applied widely in biochemical assays [10]–[14]. In our laboratory, for detecting IgM antibodies against many different viruses, µ-capture enzyme immuno assays (EIAs) have been set up by using as antigen biotinylated virus like particles (VLPs) detected by horseradish peroxidase (HRP)-conjugated streptavidin [15]–[18]. As the presence of pathogen-specific IgM antibody is applied as a marker of ongoing or recent infection, the specificity of the assay is critically important. We recently, aiming at employing as a negative control in our Merkel cell polyomavirus (MCV) IgM serology, and encountering strong reactivity in the serum of one of us authors, found the IgM to react with the biotinylated VLPs but not with the unbiotinylated ones. Prompted by this preliminary finding, the following study was initiated.
In each blot the left lane contains biotinylated BSA, and the right lane contains nonbiotinylated BSA. In lower right corner is the blot treated with the protein stain Ponceau S red.
In this study, we looked for human serum IgM antibodies reacting with biotin (Bio-IgM) and developed an indirect EIA to quantify their level and assess their seroprevalence. The adverse effects of biotin IgM on biotinylation-based immunoassays were assessed, including four inhouse virus IgM EIAs and one commercial assay. The biotin IgM and streptavidin/avidin were shown to compete for binding to biotin. The affinities of the biotin IgM were also determined.
Detection of Biotin Antibodies in Human Sera and Assessment of Seroprevalence among Adults and Children
To measure seroreactivities against biotin among the general population, an indirect IgM EIA (Bio-EIA) was set up with biotinylated BSA as antigen. The EIA OD values of 612 serum samples from 459 adults were plotted in a bar chart (Figure 1): 553 (90%) samples showed values <0.1 OD units, whereas 26 (4.2%) showed values >0.2. To confirm the EIA results, western blot (WB) was employed, as shown in Figure 2 with representative samples from each patient group. Positive WB signals were uniformly observed among the samples with EIA reactivities ≥0.3 OD units, but were rare among those with reactivities below 0.3. No signal against unbiotinylated BSA was seen with any of the samples.
(A) The distribution of reactivities measured by four inhouse virus IgM EIAs and one commercial kit; serum samples containing moderate or low levels of biotin IgM (EIA absorbance <1.200). The dotted lines indicate median absorbance values, and bars represent interquartile values. Absorbance values were analyzed using one-way ANOVA for overall comparison (P<0.0001), followed by Tukey-Kramer HSD test for pairwise comparison. #P<0.0001 compared to all the others; **P<0.0001, *P<0.05. (B) The reactivities with four serum samples containing high levels of biotin IgM (EIA absorbance >3.000). The cutoff values for positivity and negativity are: MCV 0.260 and 0.207, TSV 0.240 and 0.194, HBoV 0.167 and 0.136, B19 0.220 and 0.171, B19 Biotrin 0.342 and 0.279.
Among the 144 adults comprising students and staff, 4 (2.8%) had biotin IgM and among 316 senior individuals, 10 (3.2%), as shown in Table 1. From four of the latter, 1–3 week follow-up sera were available, and all maintained the biotin IgM signal at invariant intensity. However, the institute staff member who was initially positive with OD 2.5 in Bio-EIA lost his IgM during follow-up of six months, without IgG seroconversion. Among 678 samples from 349 children aged 1–12 years (median 2.5; mean 3.2), only one serum sample was positive. Overall, based on our assays, the seroprevalence of biotin IgM among adults of all ages was 3.0%, and was substantially lower (0.3%) among children. Of note, none of the Bio-IgM-positive samples contained IgG antibodies against biotin; neither did any of the 110 Bio-IgM-negative samples analyzed (data not shown).
Serum samples containing or lacking biotin IgM were applied in µ-capture IgM EIAs including (I) Biotinylated MCVLPs (at concentrations of 100 ng, 10 ng, 5 ng, 1 ng/100 µL), (II) Biotinylated BSA (at concentrations of 20 µg, 10 µg, 1 µg, 100 ng/100 µL) or (III) Soluble biotin (at concentrations of 20 µg, 10 µg, 1 µg, 100 ng/100 µL). The reactivities were plotted in boxplot. Five-pointed star: HRP conjugates. P: positive group comprising nine Bio-IgM positive sera. N: negative group comprising nine Bio-IgM-negative sera.
(A) In inhouse MCV IgM EIA, biotinylated MCV VLPs (10 ng/100 µL) were mixed with increasing concentrations of either unbiotinylated MCV VLPs or soluble biotin and added onto the µ-capture plates (100 µL/well). (B) The upper figures show two serum samples positive for biotin IgM; the lower figures show two serum samples positive for MCV IgM. (C and D) In Bio-EIA, four samples positive for biotin IgM were coincubated with avidin and applied to the assay. Five-pointed star: HRP conjugates.
Assessment of the Biotin Antibody Effect on IgM Antibody Assays
To assess whether the biotin IgM affects biotinylation-based antiviral IgM assays, all available Bio-IgM-positive serum samples were examined by four inhouse virus IgM EIAs of µ-capture format [MCV, Trichodysplasia-spinulosa polyomavirus (TSV), human bocavirus 1 (HBoV), parvovirus B19 (B19V)] and by one commercial kit (B19V EIA of Biotrin). In Figure 3A, 16 adults' sera with Bio-EIA ODs between 0.312 and 1.118, showed in inhouse-EIA ODs of 0.202–0.674 for MCV, of 0.045–0.338 for TSV, of 0.013–0.072 for HBoV and of 0.066–0.309 for B19V, and ODs of 0.078–0.171 in the commercial B19V EIA. In contrast, four samples (one senior sample and three of the four B19V-IgM false positive samples) with Bio-EIA reactivities exceeding OD 3, showed elevated reactivities in all assays (Figure 3B). With each of these sera, the ODs in MCV EIA exceeded those in the other EIAs. According to the cutoff values for each assay, among those 20 serum samples, the numbers of respective samples that were positive, negative, and at borderline range in each assay were 17, 3, 0 for MCV, 7, 11, 2 for TSV, 4, 16, 0 for HBoV, 7, 12, 1 for B19V (inhouse), and 1, 18, 1 for B19V (Biotrin).
In Bio-EIA, sera preincubated with increasing concentrations of either soluble biotin (5, 10, 20, 50, 75 µg/100 µL) or vitamin B5 (25, 50, 75 µg/100 µL) for 1 hr at RT, and applied into the plates (100 µL/well). The reactivities declined to undetectable levels upon serum preincubation with biotin, but not with vitamin B5.
Diluted sera were incubated with increasing concentrations of biotin over night, and then transferred into Bio-EIA plates.The amount of antibody bound to biotinylated BSA in the solid phase was measured.
Competition between IgM and Streptavidin/avidin for Binding to Biotin
Serum samples, 9 Bio-IgM positive and 9 negative, were applied in a µ-capture EIA (inhouse EIA, Figure 4), and therein tested with one of three different antigens: biotinylated MCV virus like particles (MCVLPs), biotinylated BSA and soluble biotin. In Figure 4B-I, the Bio-IgM-positive samples showed positive IgM results with biotinylated MCVLPs at a minimum amount of 5 ng/well, with a dose-dependent increase in reactivity. Correspondingly, they also yielded positive IgM results (Figure 4B-II) with biotinylated BSA at 10 µg/well or higher, but negative results with soluble biotin regardless of concentration (Figure 4B-III). By comparison, none of the Bio-IgM-negative sera showed any reactivity using as antigen biotinylated MCVLPs, biotinylated BSA or souble biotin. Competition experiments (Figure 5A and B) were performed with samples reactive with biotinylated MCVLPs in a µ-capture IgM EIA (MCV inhouse EIA). With two Bio-IgM-positive sera, the reactivities were blocked dose-dependently by increasing concentrations of soluble biotin, but not by non-biotinylated MCVLPs. Correspondingly, with two MCV-IgM-positive sera, the reactivities were blocked by MCVLPs, but not by biotin. Furthermore, as shown in Figure 5C and D, the Bio-EIA reactivities of positive sera were efficiently abolished by coincubation with avidin dose-dependently with increasing concentrations.
Affinity Determination (Half-maximal Inhibitory Concentration IC50)
The binding specificity and affinity of biotin IgM antibodies were further studied by competition experiments (Figure 6A), in which the binding of a given Bio-IgM to solid-phase biotinylated BSA was measured in the presence of homologous soluble biotin and heterologous vitamin B5 (Pantothenic acid, Sigma-Aldrich, P2250). In Figure 6B, the reactivities of six serum samples were inhibited by biotin, but not by vitamin B5. In Figure 7, with six serum samples diluted, the binding of biotin IgM to biotinylated BSA was inhibited in a dose-dependent fashion, with IC50s ranging from 2.1×10−3 to 1.7×10−4 mol/L.
We report for the first time the occurrence of IgM antibodies to biotin in humans of the general population. The biotin IgM shown here interacts non-covalently with biotin, differently from previously reported biotin-binding immunoglobulins (BBI), which were formed by biotin covalently bound to the constant region of IgM or IgG [19], [20]. For seroprevalence determination, we developed an EIA for these antibodies. The cutoff for positivity in our assay was set according to immunoblot results, whereby we found a seroprevalence of 3% among Nordic adults. Existing data on occurrence of human antibodies to vitamins are scarce except for anti-vitamin D IgG antibodies that have been observed at a low prevalence in patients with autoimmune diseases [21]. As autoantibodies do occur also in healthy people, e.g. rheumatoid factor (RF) in 1–3% of the general population [22]–[24], the possibility that the biotin IgM has an association with autoimmunity deserves to be explored in the future. On the other hand, with an essential role in metabolism, biotin can exist in the human body in various forms, such as biotinylated carboxylases and metabolites [1]. Another interesting possibility stems from the fact that biotin only accounts for half of the total avidin-binding substances in human plasma, as biotin metabolites that originate from cellular oxidation account for the rest [25], [26]. As those metabolites in molecular structure resemble biotin [1], it is possible that the biotin IgM is derived from an immune response to these metabolites, possibly due to an imbalance in biotin metabolism.
Since biotin IgM was initially found in a serum sample with false positivity in our inhouse IgM assay, we determined the susceptibility of several assays to it. We found that, serum samples with high levels of biotin IgM affected adversely all virus IgM assays, while samples containing low levels of the antibody showed low interference level in most assays. Indeed, biotin IgM was shown to be an important contributor to false positivities in virus IgM assays. Moreover, our results showed that among µ-capture IgM assays for four different viruses, the effects of biotin IgM were different. The MCV IgM EIA was more susceptible than the others. This difference may depend on the conformation of viral particles or their degree of biotinylation. Thus, at least two ways emerge to limit the effect of the biotin IgM: one is to optimize the amount of biotin in VLP trimming, and the other is to employ an appropriate borderline zone upon setting up the cutoffs according to the false reactivities derived from biotin IgM.
We did study in detail whether the different effects of biotin antibodies on IgM assays would relate with the conformations of the biotinylated antigens. As MCV VLPs were more susceptible than BSA, it is possible that ordered supramolecular assemblies are more susceptible than isolated polypeptides. Of note, biotin itself in solution never gave a positive signal. To account for this, three possibilities existed: (a) the soluble biotin did not bind to the biotin IgM captured on the plate; (b) the IgM-attachment abolished biotin's ability to bind to streptavidin; (c) IgM would bind the long spacer arm (LC) of biotinylation reagent (Sulfo-NHS-LC-Biotin) instead of the biotin itself. To find the answer, we performed competition assays with soluble biotin in MCV IgM EIA. The fact that soluble biotin displaced biotinylated MCV VLPs without causing positive results itself, proved that biotin could not bind IgM antibodies and streptavidin simultaneously and that the binding was specific for the biotin itself and not the spacer arm. As avidin is a natural constituent in human's diet and binds to biotin similarly as streptavidin does [12], we performed competition assays with avidin, which confirmed that IgM and streptavidin/avidin compete with each other for binding to biotin. As biotin is a small molecule (244Da), IgM and streptavidin/avidin probably share on biotin the same binding site. On the other hand, as avidin can prevent the intestinal absorption of biotin [3], [7], [8], it is tempting to speculate a similar function for the biotin IgM in blood. Indeed, whether the biotin IgM can prevent biotin uptake by tissues or cells is an exciting topic for further study.
We furthermore assessed the binding strength of biotin IgM by measuring IC50 [27]. The observed affinity level of 10−3–10−4 mol/L is similar to that of many natural antibodies with their respective antigens [28]–[32]. An interesting possibility is that the biotin IgM belongs to the group of germ line-encoded immunoglobulins generated without known antigenic stimulation. Most natural antibodies do belong to the IgM class and are presumed to be driven by endogenous host antigen [28], [30], [33]. As biotin has the ability to interact with host proteins [1], [2], [34], the resulting complexes might drive the production of these antibodies toward the ligand. However, the biotin IgM seemed specific, as it in our study did not react with vitamin B5, a vitamin so closely related to biotin that it can inhibit the intestinal absorption of the latter [35].
Since our study cohorts are residents in Finland, it would be worthwhile to investigate biotin antibodies in non-Finnish populations. On the other hand, as our seroprevalence data are not population-based, they do not necessarily disclose the precise prevalence of biotin IgM antibodies even among the Finnish population. Nevertheless, the finding of biotin IgM in human sera opens a new venue for research of vitamin-directed immunity. The potential effects of these antibodies on human health call for investigation. These antibodies might adversely affect the uptake of biotin from circulation, or might help in clearance of excessive biotin and its metabolites in blood.
Materials and Methods
Ethics Statement
The study protocol was reviewed and approved by the Ethics Committees of Turku and Helsinki University Hospitals. Written informed consent was obtained from all participants. If senior participant was unsure about the consent, or he/she was assessed by a geriatrician to have reduced capacity to consent, a written consent was obtained from the next of kin. If participants were children, consent was obtained from parents or guardians.
The occurrence of biotin IgM antibodies was determined in 612 serum samples from two groups of adults and 678 samples from children. The first group, referred to as "Adults" comprised 144 sera from University students and our Institute staff, and the second group, "Seniors", comprised 468 sera from 316 senior citizens (mean age 82 years, range 70–100). All senior citizens were in-patients of Turku City Hospital and participated to Virel study which aims to investigate detailed molecular virus etiology in the elderly and its links to illness severity and long-term outcome. The last group, "Children", comprised sera from 349 children at sampling age of 1–12 years (median 2.5; mean 3.2), which were previously studied for MCV and TSV primary infection [36], [37]. In addition, four serum samples observed to cause false reactivities in our inhouse assay for B19V IgM due to biotin antibodies, were also included in the present study but omitted from the calculation of seroprevalence.
Biotin IgM Indirect Enzyme Immunoassay
An indirect EIA was set up for detection of biotin IgM in human serum samples (Bio-EIA, Figure 1A). For use as antigen, bovine serum albumin (BSA) (Sigma-Aldrich, A7906) was biotinylated by using the EZ-Link Sulfo-NHS-LC-Biotinylation kit (Pierce, Rockford, IL.) in accordance with the manufacturer's instructions, and dialyzed against PBS. Strips of microtiter wells were placed in lockwell frames (Nunc-Immuno™ Modules and Frames), and coated with 200 ng/well of biotinylated BSA in PBS for 2.5 hours at 37°C. After 5 washes with PBS containing 0.05% Tween-20 (PBST), the wells were blocked in 5% BSA in PBS for 30 min at 37°C, followed by a rinse. The serum samples diluted 1∶100 in PBST were applied in duplicate (100 µl/well) for 60 min at room temperature in a rocking (400 rpm) EIA incubator. After washes with PBST, polyclonal rabbit anti-human IgM-horseradish peroxidase conjugate (DakoCytomation, Glostrup, Denmark) diluted 1∶1,000 in 3% BSA/PBST, was added to each well (100 µl), and incubated for 60 min. After washes, orthophenylene diamine and H2O2 were added, the reaction was stopped after 25 min with 0.5 M H2SO4, and the absorbances at 492 nm were recorded. For background reactivity, a BSA-EIA was set up by coating the plates with unbiotinylated BSA and performed as above.
In cutoff determination, all sera yielding OD <0.15 units in Bio-EIA were considered negative, and the samples with higher OD were re-examined by both Bio-EIA and BSA-EIA; and the net OD was obtained by subtraction of BSA-EIA background. Samples with net OD ≥0.3 were considered positive, those with net OD <0.2 were considered negative, and those with net OD between 0.2 and 0.3 were considered borderline.
Western Blot Analysis
For detection of biotin IgM by immunoblot, 1 µg/lane of biotinylated BSA (Bio-BSA) or BSA was separated by 10% SDS-PAGE and transferred onto nitrocellulose membranes (Whatman Protran BA 85), blocked in 5% BSA in Tris buffered saline containing 0.1% Tween 20 (TBST) for 30 min at room temperature (RT). The serum samples diluted 1∶100 or 1∶50 in 5% BSA/TBST were kept on the blots at 4°C overnight. After washing with TBST, a polyclonal rabbit peroxidase-conjugated anti-human IgM (DakoCytomation, Glostrup, Denmark) diluted 1∶20,000 in 5% BSA/TBST was applied for 1 hour at RT. After washes, the blots were developed using the SuperSignal West Dura Extended Substrate Detection kit (Pierce, Rockford, IL.) and exposed on X-ray films (Fuji Super RX).
IgM Enzyme Immunoassay
µ-capture EIAs were performed in analogy to our many antiviral IgM EIAs (inhouse EIAs) [15]–[18]. Briefly, serum samples diluted 1∶200 in PBST were applied in duplicate into wells of plates coated with goat anti-human IgM (Cappel/ICN Biomedicals) for 60 min at room temperature. After washes, the biotinylated antigens were applied and incubated for 45 min at 37°C. Then the plates were treated with horseradish peroxidase-conjugated streptavidin (Dako, Glostrup, Denmark) at 1∶12,000 in PBST containing 0.5% BSA for 45 min at 37°C, followed by orthophenylene diamine and H2O2 for 15 min at 37°C. In competition assay, the biotinylated antigen was mixed with a nonbiotinylated one, and added into the plates (Figure 5A).
Affinity Determination
Based on the Bio-EIA, a competition assay was set up to measure biotin IgM affinity [27], [29], [38], [39]. Briefly, the sera were diluted for an OD of 0.4 to 1.2 and incubated with increasing concentrations of soluble biotin (Sigma-Aldrich, B4501) over night, to reach equilibrium. The mixtures were then transferred into EIA plates in five replicates and the amount of antibody bound to the solid phase was measured as described above. The mean values were plotted in a graph, yielding a fitting curve by a nonlinear regression program (GraphPad Prism). The IC50 was then calculated from the curve, corresponding to affinity [27], [39].
Author Contributions
Conceived and designed the experiments: TC KH. Performed the experiments: TC LH. Analyzed the data: TC KH. Contributed reagents/materials/analysis tools: PSM LJ TJ OR MS. Wrote the paper: TC KH.
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- 36. Chen T, Hedman L, Mattila PS, Jartti T, Ruuskanen O, et al. (2011) Serological evidence of merkel cell polyomavirus primary infections in childhood. J Clin Virol 50(2) 125–129: 10.1016/j.jcv.2010.10.015.
- 37. Chen T, Mattila PS, Jartti T, Ruuskanen O, Söderlund-Venermo M, et al. (2011) Seroepidemiology of the newly found trichodysplasia spinulosa-associated polyomavirus. J Infect Dis. 10.1093/infdis/jir614.
- 38. Friguet B, Chaffotte AF, Djavadi-Ohaniance L, Goldberg ME (1985) Measurements of the true affinity constant in solution of antigen-antibody complexes by enzyme-linked immunosorbent assay. J Immunol Methods 77(2): 305–319.
- 39. Devey ME, Bleasdale-Barr KM, McLachlan SM, Bradbury J, Clark F, et al. (1989) Serial studies on the affinity and heterogeneity of human autoantibodies to thyroglobulin. Clin Exp Immunol 77(2): 191–195.


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Senior Management & Heads of Department jobs in Liverpool
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Senior Content Manager
Times Higher Education (THE), the leading global authority on university performance, is searching for a motivated and driven ...
Top job
Programme Manager – Senior Leader Degree Apprenticeship
Are you a creative, passionate and customer-focused individual with experience of teaching and managing work based ...
Top job
Dean, Jindal School of Environment and Sustainability
O.P. Jindal Global University (JGU) is an Indian University providing world class education, located in the National Capital Region ...
Top job
Director Research and Innovation Services
Liverpool John Moores University is one of the largest, most dynamic and forward-thinking universities in the UK...
Director of Student Advancement
Liverpool John Moores University is a pioneering modern civic university delivering solutions to the challenges of the 21st...
Programme Leader Industrial Automation and Control
The Department of Maritime and Mechanical Engineering is recruiting to appoint an academic leader with responsibility...
Programme Leader - Education Studies
Temporary for up to 12 months (maternity leave cover) The Faculty of Education, Health and Community has a long...
Director Research & Innovation Services
Liverpool John Moores University is one of the largest, most dynamic and forward-thinking universities in the UK...
Project Manager Clean Growth UK - Low Carbon Eco-Innovatory
Clean Growth UK will be a partnership with the University of Brighton, Liverpool John Moores University and the...
Head of Department and Chair in Communications and Media
We are seeking to appoint a Head of Department and Chair in Communication and Media. As the Head of Department, working ...
Deputy Director of Studies
Applications are invited from Consultants/Clinical Academics or Academics who have a particular interest in Medical Education and ...


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Swimming Teacher (Trainee)
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Job Type Part-Time
Company Website https://www.swimmingteacherinstitute.asia
Company Name Swimming Teacher Institute
Education Level Not Specified
Job Description
Take the splash and plunge into a highly rewarding career!
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Tech Talk
Convey Announces Executive Appointments as Company Expands Operations
With strong momentum, 300% growth in revenue run rate, and a 4X multiplier of platform users, Convey announced that it plans on doubling headcount and expanding to a larger HQ. Additionally, the shipping software startup, based in Austin, TX, announced that it would bring on four key leaders to shape future growth.Read more
The Recipe For Better Company-Building
Great companies are built by great people, and the biggest opportunity in company-building for tech is identifying the right talent who can execute on the vision and ultimately drive value for customers. Read Rob Taylor's five principles for building a value-driving team.Read more
Austin logistics startup Convey lands millions to break into new markets
Austin-based Convey, which makes software to help online retailers with shipping logistics, has raised $8.25 million to break into new markets.
Convey received the funding from Techstars Venture Capital Fund, RPM Ventures and other investors. The latest round brings the company's total funding to $15.75 million.
The company has focused on changing the way online retailers handle deliveries. It says its software makes it easier for customers to track orders and communicate with retailers, while providing retailers with information they need to solve potential problems before they happen.
Top CEOs on Culture and Leadership
How deeply an executive thinks about culture and leadership influences my assigned probabilities for a company's success. As a banker, a board member, on the executive team, and now a mentor to startups, there are several tells — some more reliable than others — and attention to leadership and culture is an important one. Check out what 6 CEOs, including Rob Taylor of Convey, have to say about how culture powers performance.Read more
Here Are The 2016 50 on Fire Winners
The 50 on Fire competition started with hundreds of nominees, which was trimmed to 150 fast-climbing people and companies. A few weeks later, a batch of 22 of Austin's most accomplished innovators analyzed them all and picked 50 of the hottest Austin tech startups and entrepreneurs.
Now, here's a list of the winners of 50 on Fire.Read more
8 Austin Tech Leaders Offer Advice for Finding a Startup Mentor
Finding a mentor can be a tricky step in the launch of a new business, and it's one that shouldn't be taken lightly. Just like personal relationships, the mentor/mentee partnership requires a balance of respect, care, honesty and trust.
A mentor at Techstars, Rob Taylor has been the first employee of seven startups over the past 20 years, filling just about every early stage role. He specializes in architecting and guiding a company's early development to minimize risk and maximize the chance of figuring it out.Read more
150+ Companies Attacking The Supply Chain & Logistics Space
Supply chain and logistics tech companies have seen funding go up significantly over the past few years as the category gains momentum. Smart money VCs and corporate venture arms are both participating in the space, and full-year run-rate projections estimate over $5B will be invested across 315 deals in 2016, representing annual highs for both deals and dollars.
The CB Insights database was used to create a market map of supply chain and logistics private technology companies attacking the space.Read more
What Would Happen If Amazon Bought eBay?
Let's be clear for a moment. If Amazon wanted to buy eBay, it would have not one, not two, but three fights on its hands before even contemplating the purchase. Still, Amazon's market capitalisation stands at almost 10 times that of eBay at the time of writing, having seen its share price grown by more than 23 times over the past decade.
We asked three industry experts about their views on the potentiality of such a deal and its impact.Read more
Commentary: UPS Shipping Price Rate Hikes
On Sept. 1, UPS announced that the UPS U.S. Ground service daily rate will increase by an average net 4.9%. Additionally, UPS Freight announced an average net 4.9% general rate increase.
The most important part of this story isn't that rates are rising. Rather, retailers need to become more effective at managing a larger network of carriers to save money, offer customers more options, and deliver a better customer experience. This focus on optimization is especially important for sellers of large-items and those items that can ship in various modes (i.e. Parcel or LTL).Read more
Silverton Partners, Capital Factory pump $4.5M into Austin software maker
Convey Inc., a shipping rate comparison provider, announced that it has completed a $4.5 million Series A round of funding. Investors included Silverton Partners, Techstars Ventures, the Capital Factory and Corsa Ventures. The Austin company that changed its name from Pivot Freight Inc. plans to invest the capital in its engineering work.
In early 2015, Convey enlisted as its CEO Rob Taylor, who was previously one of the founding executives of California-based TrueCar Inc. (Nasdaq: TRUE) and Austin-based BlackLocus Inc., which was acquired by Home Depot Inc. (NYSE: HD) in late 2012.Read more


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Jobs in Satellite Beach, FL
Ashley Furniture06 Aug
Professional Furniture Sales
Job Title: HomeStore Retail Sales Associate Organizational Unit: Ashley Furniture HomeStore -> Retail Operations...
Satellite Beach, FL
SenText17 Aug
Merchant Marketing Sales Agents
Job DescriptionNEED SOME FAST CASH? WRITE A DEAL TODAY AND GET PAID UP TO $748 BY DINNER TONIGHT!Build Your Own Mobile...
Satellite Beach, FL
Dollar General Corporation14 Aug
STORE MANAGER in SATELLITE BEACH, FL
Overview:Dollar General (NYSE: DG) is a Fortune 200 company with more than 15,000 retail locations in 44 states, 15...
Satellite Beach, FL
Hair Cuttery14 Aug
Hair Stylist
DescriptionSalary: $35,000 - $59,000 / year (Base salary plus tips)As a Hair Stylist for Hair Cuttery you will use your...
Satellite Beach, FL
Briel & Associates Land Surveyor...08 Aug
Experienced Land Surveyor Party Chief
Job DescriptionWe are a Brevard County Land Surveying Company seeking EXPERIENCED Land Survey PARTY CHIEF for a Full...
Satellite Beach, FL
JMJ FAMILY PRACTICE INC29 Jul
Medical Assistant
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Retail Store Associate
Description : Job DescriptionRetail Store Associates play a meaningful role within the CVS Health family. At CVS Health...
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Smile Brands Inc03 Jul
Dental Assistant
Dental AssistantSatellite Beach, FLZip Code: 32937Job Family: Dental AssistantBrand Name: Bright Now! DentalOffice...
Satellite Beach, FL
Integrative Humancare15 Aug
Licensed Massage Therapist
Licensed Massage Therapist opportunity for Integrative Humancare, an established, respected Massage and growing Wellness...
Melbourne, FL
Metlife14 Aug
Associate Director, Agricultural Investments
MetLife Investments is a premier asset manager with breadth across various asset sectors and global markets, and strong...
Melbourne, FL


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flipb Software 4.2.1
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WebMatrix is a free web development tool from Microsoft that includes everything you need for website development. Start from open source web applications, built-in web templates or just start writing code yourself. It's ...
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... in the animated GIF format for use on web pages and the AVI format (in Pivot Animator 3 and later). Pivot provides a simple, easy to use interface with ...
Photo Slideshow Creator 4.25
Photo Slideshow Creator allows you to create cool slideshows of your ... with music, animated transitions between slides and fancy designs. Using the program, you can create a great ... the Internet or on mobile devices. Photo Slideshow Creator makes it easy and quick to create a ...
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Quick 'n Easy Web Builder 5.4.2
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1st Autorun Express 3.2
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Web Page Maker 3.22
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UvScreenCamera 184.108.40.206
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Jutoh 2.80.0
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BotEngine 1.0
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XP Artistic Icons 4.0
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