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Anova Marks 1,500th Prostate Cancer Patient Treated with the Accuray CyberKnife® System
- Accuray and Anova Cancer Care Remember Prostate Patients this Father's Day
- CyberKnife System Provides Grandfathers, Fathers, Brothers, Uncles and Sons with Precise Radiation Treatments in Fewer Treatment Sessions than Conventional Radiation Therapy
Jun 16, 2017, 07:30 ET
SUNNYVALE, Calif., June 16, 2017 /PRNewswire/ -- In honor of Father's Day, Accuray Incorporated (NASDAQ: ARAY) joins Anova Cancer Care in recognizing the center's commitment to providing the most advanced treatments available to men with prostate cancer, many of whom are fathers. The center, located in Lone Tree, Colorado, has treated its 1,500th prostate cancer patient using the CyberKnife® System—an advanced radiation therapy device designed to deliver stereotactic body radiation therapy (SBRT). SBRT is a highly precise form of radiation therapy which maximizes dose to the tumor and minimizes dose to surrounding healthy tissue.
According to the American Cancer Society, more than 161,000 men will be diagnosed with prostate cancer in 2017. It's important for men to take action to improve their health, including learning about medical conditions which specifically impact them, like prostate cancer, and options for managing the disease. This Father's Day, June 18, 2017, men and their families can learn about prostate cancer at rallies held by:
- ZERO – The End of Prostate Cancer
- The Prostate Health Education Network (PHEN)
The prostate gland can move unpredictably throughout the course of treatment because of normal patient bodily functions – such as filling of the bladder, gas in the bowel, or even patient movement during the procedure – making the ability to track, detect and correct for motion critically important. Unlike any other radiation treatment, the CyberKnife System continually tracks and automatically corrects the beam for movement of the prostate in real-time throughout the entire treatment session. Published data reinforce that CyberKnife prostate SBRT is effective, with minimal side effects and impact on quality of life during and after treatment, important considerations for men diagnosed with prostate cancer.
Additional Information About CyberKnife Prostate SBRT
- Treatment with the CyberKnife System is non-invasive (no surgery involved) and is completed in 4-5 treatment sessions, over the course of one or two weeks. Conventional radiation therapy typically requires 30-40 treatment sessions, over the course of several weeks;
- CyberKnife treatment accuracy is achievable without the need for invasive rectal balloons or spacers to spare the rectal wall;
- The system can precisely target tumors and as a result, may provide a safe radiation treatment option even for some previously irradiated patients;
- SBRT increases patient satisfaction by reducing discomfort and scheduling inconvenience versus conventional radiation treatments.
Tim B.: "When I was diagnosed with prostate cancer I immediately imagined the worst. I have several grandchildren who I want to spend time with as they grow up. I discussed treatment options with my urologist and radiation oncologist and together, we decided that treatment with the CyberKnife® System was the best option for me. I liked that the technology is really precise so the radiation would be targeting my prostate and my risk for side effects would be minimized. And the convenience of only needing five days of treatment was a real bonus."
Lee K. McNeely, M.D., Radiation Oncologist at Anova Cancer Care: "Anova Cancer Care is a busy radiation therapy center, so it's important that we have technology we trust to provide our patients with the best possible treatment option. We use the CyberKnife for prostate cancer because it offers distinct advantages when treating this disease, most notably it's the only system available with the ability to track movement of the prostate and automatically adjust the radiation beam during treatment. The CyberKnife System's accuracy is sub-millimeter, enabling my colleagues and me to treat the disease with unparalleled preservation of healthy tissue and provide substantial quality of life and lifestyle benefits to patients."
Stanley H. Galansky, MD, FACS, FAAP, Urologist at Urology Associates: "Urology Associates offers a range of treatment options for prostate cancer, including the CyberKnife, which we administer on an out-patient basis at Anova Cancer Care. We partnered with Anova to offer our patients this option because we believe it effectively manages the cancer with considerably fewer side effects than traditional cancer treatments. With the CyberKnife System, we treat patients in 5 sessions and it's less expensive for the patient and their insurance company. Most importantly, the system has provided quality outcomes for our patients."
Joshua H. Levine, President and Chief Executive Officer of Accuray: "We commend Dr. McNeely and the Anova Cancer Care team on their dedication to improving the lives of their patients. We appreciate our continued partnership and are honored they have selected the CyberKnife technology to treat so many of their low- to intermediate-risk prostate cancer patients. The CyberKnife System is continuing to demonstrate its position as the radiation treatment of choice for more and more physicians and patients. It offers a convenient, non-invasive approach which precisely targets the prostate cancer with minimal disruption to patients' daily lives."
Important Safety Information
For Important Safety Information please refer to http://www.accuray.com/safety-statement
About Accuray
Accuray Incorporated (Nasdaq: ARAY) is a radiation oncology company that develops, manufactures and sells precise, innovative treatment solutions that set the standard of care with the aim of helping patients live longer, better lives. The company's leading-edge technologies deliver the full range of radiation therapy and radiosurgery treatments. For more information, please visit www.accuray.com
Safe Harbor Statement
Statements made in this press release that are not statements of historical fact are forward-looking statements and are subject to the "safe harbor" provisions of the Private Securities Litigation Reform Act of 1995. Forward-looking statements in this press release relate, but are not limited, to clinical applications, clinical results, patient experiences and outcomes, and Accuray's leadership position in radiation oncology innovation and technologies. Forward-looking statements are subject to risks and uncertainties that could cause actual results to differ materially from expectations, including but not limited to the risks detailed from time to time under the heading "Risk Factors" in the company's report on Form 10-K, filed on August 24, 2016, the company's reports on Form 10-Q, filed on November 1, 2016, February 3, 2017, and May 5, 2017, and as updated periodically with the company's other filings with the SEC.
Forward-looking statements speak only as of the date the statements are made and are based on information available to Accuray at the time those statements are made and/or management's good faith belief as of that time with respect to future events. The company assumes no obligation to update forward-looking statements to reflect actual performance or results, changes in assumptions or changes in other factors affecting forward-looking information, except to the extent required by applicable securities laws. Accordingly, investors should not put undue reliance on any forward-looking statements.
Beth Kaplan
Public Relations Director, Accuray
+1 (408) 789-4426
[email protected]
Jane Hauser
+1 (781) 684-0770
[email protected]
SOURCE Accuray Incorporated


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Zatecka N°14 , Old Town - Prague 1
up to 5 people in 3 rooms, 1 bathroom, 2 toilets
Show on map> Show daily rates>
The two-bedroom, one-and-a-half bathroom Rudolfinum Apartment, complete with a balcony, is situated in a gorgeous Art Nouveau apartment building with 24-hour reception and a lift, and is located in Prague's high-end sought after shopping and dining area of Josefov (Jewish quarter). The holiday apartment features free Wi-Fi Internet, satellite TV with Internet access, washer/dryer, dishwasher and more. The price includes weekly cleaning, full linen and towels. The price is for 4 persons; extra person is EUR 15 per night.
This classic Prague apartment has two bedrooms (one of which boasts a balcony, while the other features a traditional ceramic stove), a living room with sofa / entertainment area, fully fitted kitchen and dining area; a full bathroom with bathtub, wash basin and toilet, and a WC (with 2nd toilet, sink). The apartment has original Art Nouveau ceiling plasterwork and hardware (window latches, locks, etc.) throughout, as well as parquet and tile flooring. The space has been decorated and furnished with upmost style and comfort in mind. You will need to walk up a short flight of stairs from the lift landing to get to the apartment.
The apartment building is situated directly in the center of Prague's Old Town and Jewish quarter (Josefov). Set on the corner of two bustling streets across from the Old Jewish Cemetery and the Pinkas Synagogue (which now functions as a memorial for the Holocaust victims from Czech lands), this is a most memorable address. The neighborhood can be characterized by its abundance of high end fashion boutiques, antique and jewelry shops, and fine dining establishments (offering an array of cuisines – Czech, Kosher, international). A variety of nightlife and entertainment options are available as well. The Vltava River, Rudolfinum concert hall & art gallery, and the Museum of Decorative Arts are in the immediate vicinity, while Old Town Square, Charles Bridge, Lesser Town, and New Town are within comfortable walking distance.
Zatecka N°14
Žatecká 14
Praha 1 - Old Town
110 00

subset: commoncrawl


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Here is another post from the archives.
There are so many social media outlets today that it can be challenging to keep them all straight. While I do use most mainstream platforms I'm grateful that social media wasn't a part of my life till after college. Although I have to admit there are some perks to being this connected. Facebook allows me to have a relationship with cousins and distant relatives I've yet to meet in person, Twitter keeps me up to date on great events and contests I wouldn't have known about otherwise, and I do enjoy seeing photos others capture on Instagram. I downloaded Snapchat when it first came out but quickly gave up on that. It was one platform too many for me.
My favorite platform by far is Pinterest. It has become as much a part of my weekend routine as coffee and the Sunday New York Times. Pinterest is where I find inspiration for ensembles I want to add to my wardrobe, recipes I'm dying to try, and DIY projects for a rainy Saturday.
One of the fun things about this blog has been connecting with subscribers via Pinterest. I love seeing what gems others find and it's interesting to see what pins of mine others are drawn to. I mentioned that Pinterest is part of my weekend routine. Throughout the week I "like" pins that seem interesting to me and on the weekend I organize those pins onto my different boards. I create boards for inspiration when I'm planning out parties.
Are you an avid pinner? Please share the link to your Pinterest profile in the comments so we can all follow.


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Seeing more blue light and less red light most of the time, I am looking for the red light in the atmosphere to balance my perception of the spectrum. I notice if I see less red light, that I feel less energy in my legs and feet than I usually do. When I perceive more red light or I find I can wear glasses to either block blue or enhance red, I suddenly feel more grounded and more secure.
Tag: highly sensitive children
Video Snippet – Seeing Outside of the Visible Spectrum – Thinking Outside of the Box!
Some days the light feels very strange. The roofs I see from my window stand out in their darkness and other black markings on birds appear differently to how they usually do. On those day the light also seems very bright. This might happen on sunny days in the middle of summer but can happen when there is lots of white cloud at other times of the year too. I can find myself squinting inside, even on a January day.
I Bought Myself a Spectrometer!
Hi, I have been making a lot of videos about how I perceive light and colour in a different way to those around me. I have often wondered if I am 'seeing' differently or 'processing' differently so I bought myself a spectrometer!
My Discoveries & Thoughts
I perceive light differently I constantly perceive subtle changes in the colour spectrum in the atmosphere. For instance just looking at the sky I can perceive when there is an increase in violet light or decrease in red light. I have been testing my observations with my spectrometer readings, and I find that how I perceive the light does not always tally with what my spectrometer picks up.
My brain sometimes processes the relative intensity of wave lengths in a way that is atypical. This means that I might perceive some wave lengths as less intense or more intense than I see on my spectrometer.
(incidentally my colour sensitivity is so extreme that I can't look at the intensity of a colour spectrum so my husband removed the colour and sent me a black and white line graph for my observation.)
I feel the change in the light as much as I perceive it visually. I know if there is more violet light because I will feel more intuitive or suddenly have a desire to eat crunchy food. I know if there is more red light because I will feel more grounded and have a desire to eat softer foods.
I live by the light meaning that from when I get up the morning I have half an eye out to check what the light is doing. How I see the light affects all my choices of the day – what I wear, what I eat, what I do and how I relate to myself and other people. When I perceive more violet light I am more creative and intuitive. When I perceive more red light I am more into my home projects. When I perceive more yellow light I enjoy relationship with other people more.
My imbalanced way of seeing the light moves me and shapes me. It sharpens my intellect, helps me tune into my emotions and heightens my creativity and intuition.
Living by the Colours I See in the Light – The Joys and Challenges of Having Atypical Spectral Sensitivity
Video Snippet: Stormy Days – There is a Lot of Energy Out There!
On stormy days I feel intense and often agitated. I struggle to think clearly and in the end I just have to get on with something and the more intense it is the better!
Video Snippet – My Reaction to Green can be Extreme!
My perception of the light can make some colours seem too bright on some days. Here I share my experience of going to a park on a particular June day after it had rained.
Video Snippet – The Green Light of Digestion!
Perceiving more green light affects my digestion of who I am, my life, love , praise . . . anything that I need to process and digest.
Video Snippet – Sometimes the Colours Look Wrong to My Brain!
Every day the colours change as the light changes. Sometimes colours look 'wrong'. Sometimes they look 'right.' This greatly impacts how I live my daily life.
Video Snippet – When the Violet Light Drops Does Your Mood Drop Too?
I am like a mountaineer enjoying immensely being on the mountain tops. This is what it feels like to perceive more violet light. I am full of buzz, excitement and ideas. When it drops, though, I have to find a more open hearted way to connect with my happiness.
Video Snippet – The Bright Cloud of Spontaneity and Freedom
There are many kinds of cloud and bright storm clouds make me feel so hemmed in that I just crave freedom.
Video Snippet – Being the Pusher!
In certain types of cloud I feel as if my energy is being compacted and the only answer is to be the pusher – to push through with some project or idea.


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Hunger, Climate Change and Immigration
6:30 pm Sunday, February 22 Hunger, Climate Change and Immigration: The Nueva Frontera program in Honduras
6:30 pm Sunday, February 22 Hunger, Climate Change and Immigration: The Nueva Frontera program in Honduras
Written by Jerry Mogensen At First Christian Church, we are all about love and justice. As many of you know, we are founding members of what is now Central Iowa Shelter Service located at 1420 Mulberry Street. This shelter houses… More
Written by Ben Allaway With today's scripture being the famous Song of Mary, also known as the Magnificat, where Mary praises God in response to the revelation that she will bear the Messiah, it is a good opportunity to share a… More
First Christian Church's first Conference for Faith & Learning is coming to on October 17-19 and an exciting weekend of activities is planned! Focused on the theme of covenant, participants will learn about biblical and historical covenants, how covenant impacts our lives… More
This year's theme, #Reconciliation Generation: School Yards Not Prison Yards, is undergirded by Psalm 78 which reminds us of God's steadfast blessings in our past and present, and God's faithfulness toward restorative justice. God's unfathomable grace toward each of us informs our courage to impact future generations for Christ's shalom. As the Christian Church (Disciples of Christ,) we are committed to addressing racism as a sin and restoring all of God's children to the fullness of their humanity. More
A Fun Weekend of Learning, Worship, Fellowship and Service is Coming to First Christian Church! The Conferences for Faith and Learning each focus on a theme to explore, discuss and practice. This month's conference theme is: Covenant. RSVP by calling… More
On Saturday, September 20th, First Christian Church's Sunday Learning Hour Class, People, Inc., took its annual turn serving the evening meal at Central Iowa Shelter and Services in downtown Des Moines. An estimated 250 persons were served tater tot casserole… More
We wish to thank so many who responded when Justice in Action was activated in response to the unaccompanied children flocking to our borders for asylum and help from the dangerous and deplorable conditions in their home countries. Our first… More
The humanitarian crisis of the 50,000+ unaccompanied migrant children being processed through detention centers across the country is, by now well published. "Who Are the Kids of the Migrant Crisis?" from NPR "Faith Leaders Have Right Response to Immigration… More
This is my 5th summer taking First Christian youth on a summer mission trip. This year our high school youth are in Kansas City, MO. Each trip has its own significant memories, each trip strengthens connections between our young people,… More


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Hello Commanders,
WD Division Leaderboards will be introduced later today for future World Domination events. We've prepared a short FAQ for your inquiring minds. Please feel free to leave comments and questions below. Thanks.
What is the Division Leaderboard?
You will now see a third tab for leaderboards in World Domination events. This leaderboard will be comprised of a competitive mix of Factions based on past event performance.
Where can I find my Division Leaderboard?
You will notice a third tab available on the World Domination Leaders tab screen. This tab will contain your division. You will not be able to see the other division leaderboards.
Does this mean there are more prizes available?
Yes, with this introduction of a new leaderboard, we have added an additional set of prizing. All divisions will consist of an identical prize pool. You will see an additional "Divisions" tab on the Rewards screen.


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Partner: Toronto Rehabilitation Institute – University Health Network
Re: Traumatic brain injury in the workplace: Innovations for prevention
Date: August, 2015
Description of the work:
We are looking to collaborate with a creative, socially-minded animator or animation company with an interest in social change. As a group of clinical and health services researchers, we are looking to use animation as a form of 'knowledge translation' which will help make our research on work-related traumatic brain injury accessible to members of the community.
We have two complimentary animation projects. One is immediate: a 2-3 minute information/'explainer' animation about traumatic brain injury/concussion geared towards adults and the work place (as opposed to sport or children). The second is a longer-term project: we are looking to make a series of short animation vignettes based on the experiences of individuals who experienced a brain injury at work. Likely, this second project will be more nuanced and artful than the first. For the time being, we're focusing on the Explainer Video, but there is the possibility to collaborate on the Vignettes in the future.
Who you are:
We are looking for someone who is an exceptional animator, with the ability to work in a cross-disciplinary team. Additionally, you must be attentive to the needs of people with disabilities and accessibility standards.
Who we are:
You will be collaborating with a team of international leaders in brain injury research at the Toronto Rehabilitation Institute – University Health Network, and arts-based knowledge translation specialist, Julia Gray.
The Traumatic brain injury in the workplace: Innovations for prevention research study will improve our understanding of occupational brain injury, and inform future initiatives designed to improve workplace safety by helping prevent work-related brain injury. Please visit the Acquired Brain Injury Research Lab website for more details: http://www.abiresearch.utoronto.ca/
Julia Gray is a writer, theatre director and arts-based researcher in health. She has collaborated with health and social scientists for 10 years and has developed several artistic projects based on research as a way to engage members of the community. Julia is currently collaborating with the research team to develop the explainer video script and is spearheading the coordination of the project. www.possiblearts.ca
What we need:
Please submit a quote for the 2-3 minute explainer video ONLY, including details about process (e.g. script consultation/feedback, sketches, story board, animation, voiceover/sound/music) and costs. Please also submit a short biography, 2 references and a link to portfolio. Please make submissions by October 9, 2015 at 5pm to Julia Gray at email@example.com


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How to Make It
Step 1
Preheat grill to high. Brush cut sides of baguette with 1 Tbsp. olive oil and sprinkle with salt. Grill bread, cut sides down, until lightly charred, about 1 minute. Let cool and cut into 1-inch pieces.
Step 2
Brush chicken all over with 1 Tbsp. olive oil and sprinkle with salt. Grill, turning once, until cooked through, 6 to 10 minutes. Let cool slightly.
Step 3
Combine tomatoes, cucumber, olives and bread in a large bowl. Whisk together remaining 6 Tbsp. oil, lemon juice, yogurt and garlic in a small bowl. Season with salt and pepper. Toss dressing and basil with bread mixture to coat. Divide among 4 dinner plates.
Step 4
Thinly slice chicken and place on top of salad. Serve immediately.


text: Travel Grant Application No Share Share this page on Twitter Share this page on Facebook Share this page on LinkedIn Print this page Email this page Page ContentI wish to apply for a Travel Grant to be used to offset my institution's expenses incurred for Institute registration, food, lodging, and travel for the Presidents Institute, January 4–7, 2020, in Marco Island, Florida.This application must be submitted no later than 5:00 p.m. Eastern time on Friday, November 1, 2019. For proper tracking of applications and for security reasons, accessing the application form requires authentication. To review the application form and to submit an application, please use the Sign In link at the top of the page to sign in with your username (institutional email address) and CIC password. If you do not know your CIC password, you can retrieve your password online (If you do not have an account, you will be prompted to create one).


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Molluscan smooth muscle can maintain tension over extended periods with little energy expenditure, a process termed catch. Catch is thought to be regulated by phosphorylation of a thick filament protein, twitchin, and involves two phosphorylation sites, D1 and D2, close to the N and C termini, respectively. This study was initiated to investigate the role of the D2 site and its phosphorylation in the catch mechanism. A peptide was constructed containing the D2 site and flanking immunoglobulin (Ig) motifs. It was shown that the dephosphorylated peptide, but not the phosphorylated form, bound to both actin and myosin. The binding site on actin was within the sequence L10 to P29. This region also binds to loop 2 of the myosin head. The dephosphorylated peptide linked myosin and F-actin and formed a trimeric complex. Electron microscopy revealed that twitchin is distributed on the surface of the thick filament with an axial periodicity of 36.25 nm and it is suggested that the D2 site aligns with the myosin heads. It is proposed that the complex formed with the dephosphorylated D2 site of twitchin, F-actin and myosin represents a component of the mechanical linkage in catch.
Molluscan smooth muscles, such as the bivalve adductor muscle and mussel Mytilus anterior byssus retractor muscle (ABRM) are capable of maintaining tension for extended periods with little energy consumption. This phenomenon is called catch. Catch muscles contract by stimulation with acetylcholine and the resulting increase in intracellular [Ca2+]. Subsequent removal of acetylcholine results in the maintenance of tension even at low intracellular [Ca2+] (Ishii et al., 1989; Twarog, 1976). Secretion of serotonin abolishes the catch state by increasing intracellular [cAMP] and activation of cAMP-dependent protein kinase (PKA). These various phases (Watabe and Hartshorne, 1990) can be reproduced in skinned catch muscle fibers, i.e. an increase in [Ca2+] initiates contraction, a decrease in [Ca2+] promotes catch and addition of cAMP terminates the catch state.
Since the discovery of catch there has been considerable interest in defining the molecular basis responsible for force maintenance with very low energy expenditure. Early ideas suggested that two types of linkages were formed, the catch linkage consisting of paramyosin–paramyosin interactions and the contractile linkages of actin and myosin (Ruegg, 2001; Watabe and Hartshorne, 1990). In this scheme, phosphorylation of paramyosin by PKA would reduce or eliminate the paramyosin association and in the presence of Ca2+ allow cross-bridge cycling. Subsequently, the paramyosin hypothesis became less popular and catch was suggested to reflect attached non-cycling cross-bridge–actin interactions, the so-called `linkage' hypothesis (Lowy et al., 1964). It was suggested that phosphorylation of the myosin rod was the critical regulatory step (Castellani and Cohen, 1987). It was found subsequently that the only protein phosphorylated by PKA in ABRM (catch) fibers was the high molecular mass protein, twitchin (Siegman et al., 1997). These and subsequent studies (Siegman et al., 1998; Funabara et al., 2001a) proposed that twitchin is an integral component of the catch mechanism and is a phosphorylation-dependent regulator of the catch state. Dephosphorylated twitchin promotes catch and phosphorylation of twitchin by PKA releases catch. From in vitro studies, involving isolated thick and thin filaments under relaxing conditions, it was further suggested that the only components necessary for catch were actin, myosin and twitchin (Yamada et al., 2001). As a working hypothesis it was assumed that catch reflected attached non-cycling cross-bridges that bear the mechanical load and whose formation is regulated by phosphorylation of twitchin (Butler et al., 2001). But, other observations made it necessary to reconsider a basic tenet of the `linkage' hypothesis. Galler and coworkers found that catch depends on cAMP- and pH-sensitive linkages but not on the cross-bridge–actin interactions (Galler et al., 2005). It has also been suggested that the mechanical link in catch could be the interaction between dephosphorylated twitchin and actin, where phosphorylation of twitchin abolishes interaction (Shelud'ko et al., 2004). Binding of synthetic myosin filaments to F-actin filaments and dephosphorylated twitchin has been shown, but not direct binding of twitchin to actin (Yamada et al., 2001). The idea that alternative links may be involved was further developed and although regulation of catch by twitchin phosphorylation is accepted there is considerable evidence to support the idea that the mechanical links in catch are not attached cross-bridges (Andruchova et al., 2005; Hopflinger et al., 2006; Butler et al., 2006).
The Mytilus ABRM twitchin is composed of a single polypeptide of 530 kDa containing multiple repeats of immunoglobulin (Ig) and fibronectin type III motifs and a single kinase domain (Funabara et al., 2003) and is very similar to twitchin and twitchin-related proteins from other species (Funabara et al., 2005). In vitro phosphorylation assays with PKA indicate that ABRM twitchin incorporates 3 mol phosphates mol–1 twitchin (Funabara et al., 2001a). Two of the three phosphorylation sites are referred to as D1 and D2 and are located at S1075 and S4316 near the N and C termini, respectively. It was suggested that both sites are involved in the regulation of catch contraction in vivo (Funabara et al., 2003).
In light of the suggestion that the mechanical links in catch involve interactions other than the force-generating actin–myosin complex, this study was initiated to examine interactions with twitchin, with an emphasis on effects mediated by phosphorylation at the D2 site. Because of the high mass of twitchin (and complexity in binding assays) a D2 peptide (composed of the D2 site plus two Ig motifs) was used. Important observations were that the dephosphorylated D2 peptide binds to myosin and F-actin and promoted complex formation between myosin and F-actin. In addition the localization of twitchin on myofilaments of ABRM was examined. These cumulative data are interpreted to indicate that the D2 site of twitchin mediates interaction between myosin and F-actin.
Materials and methods
Protein preparation
The twitchin D2 region was expressed in bacteria as TWD2-S and consists of the 21st to 22nd Ig motifs from the N terminus including the D2 region (Fig. 1A). The cDNA fragment coding for TWD2-S was amplified by PCR using a primer set of 5′-CCCTCGAGCGCAAAGAAGCAGCTCCCAG-3′ and 5′-GGCTCGAGTCAGTCAAAGTCATGAGCTCGGTC-3′ and inserted into an expression vector pET-15b (Novagen, Darmstadt, Germany) after digestion with XhoI. The mutants where the D2 phosphorylatable serine (S4316 in the full sequence) was replaced by alanine (TWD2-A) or aspartic acid (TWD2-D) were constructed by PCR (Ito et al., 1991). ABRM actomyosin was prepared as previously reported from Mytilus galloprovincialis Lamarck (Funabara et al., 2001b). Chicken skeletal actomyosin was prepared by the same method as that for ABRM actomyosin. Myosin was prepared from chicken fast skeletal and scallop Patinopecten yessoensis Jay striated muscles (Stafford et al., 1979). Paramyosin was isolated from ABRM (Watabe et al., 1989). Actin was prepared from acetone powder of chicken fast skeletal muscle (Mommaerts, 1951). Twitchin was prepared as described (Funabara et al., 2001a).
Mg2+-ATPase assay
ATPase assays were carried out in 20 mmol l–1 3-(N-morpholino)propanesulfonic acid (MOPS)-NaOH (pH 7.4) containing 30 mmol l–1 KCl, 2 mmol l–1 MgCl2, 1 mmol l–1 ATP, 80 μg ml–1 actomyosin, 16 μg ml–1 D2 peptide mutant and 0.1 mmol l–1 CaCl2 or 1 mmol l–1 ethyleneglycol bis(2-aminoethylether)tetraacetic acid (EGTA). ATPase of chicken fast skeletal actomyosin was also assayed in the presence of 80 μg ml–1 ABRM paramyosin. Liberated inorganic phosphate was determined by the Malachite Green method (Kodama et al., 1986).
Solid-phase binding assay
Solid-phase binding assay was carried out according to published methods (Weitkamp et al., 1998) with some modifications. In this assay, washing, blocking reaction, dilution of samples and detection were carried out using reagents included in Protein Detector ELISA kit (KPL, Gaithersburg, MD, USA). Wells of an enzyme-linked immunosorbent assay (ELISA) microplate were coated with 100 μl of 10 μg ml–1 scallop striated adductor myosin, ABRM paramyosin or chicken fast skeletal F-actin in 1× Coating Solution included in the kit for 1 h at room temperature. After blocking, 100 μl of 10 μg ml–1 TWD2-S or thiophosphorylated TWD2-S were hybridized in 1× Coating Solution for 1 h at room temperature with proteins coated on the plate. Anti-6xHis and anti-mouse IgG antibodies conjugated with alkaline phosphatase were used, respectively, as the primary and secondary antibodies. The reaction was traced by measuring absorbance at 630 nm after the substrate solution included in the kit was added to wells.
Cosedimentation binding assay
30 μg ml–1 scallop striated adductor myosin, 300 μg ml–1 chicken fast skeletal F-actin and 10 μg ml–1 TWD2-S were mixed in 20 mmol l–1 MOPS-NaOH (pH 7.4), containing 30 mmol l–1 KCl, 4 mmol l–1 MgCl2, 1 mmol l–1 ATP and 1 mmol l–1 EGTA, on ice for 30 min and centrifuged at 3000 g. The supernatants and precipitates obtained were subjected to SDS-PAGE.
Identification of twitchin binding region of actin
Chicken fast skeletal actin was extensively digested by trypsin in 20 mmol l–1 Tris-maleate (pH 7.0) containing 50 mmol l–1 KCl and 10 mmol l–1 CaCl2 for 24 h at 37°C. The digests were subjected to reverse-phase high-performance liquid chromatography with a TSKgel ODS-80T column (4.6 mm×15 cm) (Tosoh, Tokyo, Japan) and adsorbed peptides were eluted with 0%–40% acetonitrile linear gradient. The isolated peptides were subjected to the solid-phase binding assay using TWD2-S as a probe. The peptide bound to TWD2-S was sequenced using the Applied Biosystems Procise 492 HT protein sequencer (Applied Biosystems, Foster City, CA, USA).
Competitive binding assay
A peptide AGFAGDDAP synthesized based on the sequence A21–P29 of chicken fast skeletal actin was subjected to competition binding assay for TWD2-S according to the method of the solid-phase binding assay using the same ELISA kit. Wells of a microplate were coated with 100 μl of 50 μg ml–1 chicken fast skeletal F-actin in 1× Coating Solution. After washing and blocking, 100 μl of 5 μg ml–1 TWD2-S was hybridized with the adsorbed F-actin for 1 h at room temperature. After washing, 100 μl of 0, 50, 100 and 150 μg ml–1 synthetic peptide were added to the wells and hybridized in 1× BSA Diluent/Blocking Solution included in the kit for 1 h at room temperature. Detection was performed as described in the solid-phase binding assay. Assays were done in the absence of added Ca2+.
Production of anti-TWD2-S antibody and its specificity
Anti-twitchin D2 antibody used for electron microscopy was raised against TWD2-S in rabbit. The specificity of the antibody to twitchin was confirmed by immunoblotting. To confirm whether anti-TWD2-S antibody used in the present study reacts only to twitchin in ultrathin sections and isolated thick filaments, immunoblotting analysis was carried out. Myofibrillar proteins were electroblotted onto a polyvinylidene difluoride (PVDF) membrane after separation on SDS-PAGE. The membrane was hybridized with anti-TWD2-S antibody for 1 h at room temperature and reacted with anti-rabbit IgG antibody conjugated with horseradish peroxidase as the secondary antibody after washing. Detection was carried out using 0.2 mg ml–1 3,3′-diaminobenzidine tetrahydrochloride and 0.005% H2O2.
To examine the reactivity of the antibody to twitchin and its phosphorylated form, immunoblotting analysis and ELISA using TWD2-S and its phosphorylated form were performed. TWD2-S was phosphorylated as described previously (Funabara et al., 2001a). Immunoblotting was performed as described above except for twitchin peptides. ELISA was carried out using the same kit used in the solid-phase binding assay in the present study. Wells of a microplate were coated with 10, 20 and 30 μg ml–1 twitchin peptide, TWD2-S or its phosphorylated form, and anti-TWD2-S antibody was added to the wells after blocking. Anti-rabbit IgG antibody conjugated with alkaline phosphatase was used as the secondary antibody. All solutions for ELISA were included in the kit.
Electron microscopy
Ultrathin sections for electron microscopy were prepared from ABRM fibers fixed with 4% paraformaldehyde (pH 7.2) in active contraction raised by 1 mmol l–1 acetylcholine, catch state provoked by subsequent washing with artificial seawater, and relaxation after treatment with 1 μmol l–1 serotonin. These contraction–relaxation stages were monitored by tension measurement. Fibers showed maximum tension at 50 s after treatment with 1 mmol l–1 acetylcholine (at 25°C), and at this point were fixed with paraformaldehyde and represented fibers in the active state of contraction. Paraformaldehyde-fixed fibers were dehydrated with ethanol and treated with Lowicryl K4M resin. Ultrathin sections of about 80 nm were placed on Ni-150 meshes covered with collodion membranes, treated with the anti-twitchin D2 primary antibody at 0.1 mg ml–1 for 1 h, and then with AuroProbe EM goat anti-rabbit IgG conjugated with colloid gold as the secondary antibody for 30 min each on ice. The ultrathin sections thus prepared were stained with 1% uranyl acetate and 1% lead citrate each for 10 min at room temperature, and observed using a JEOL JEM 2000EX transmission electron microscope.
ABRM thick filaments for electron microscopy were prepared according to Nonomura's method (Nonomura, 1974). Briefly, ABRM relaxed by 1 μmol l–1 serotonin was treated with 0.05% saponin and the resulting skinned fibers were homogenized on ice in an appropriate volume of 10 mmol l–1 phosphate buffer (pH 6.8) containing 0.1 mol l–1 KCl, 5 mmol l–1 MgCl2 and 10 mmol l–1 ATP. The homogenate containing thick and thin filaments was mixed with the anti-twitchin D2 antibody at 0.1 mg ml–1 for 5 min on ice, followed by reaction with the above secondary antibody conjugated with colloid gold for 5 min on ice. The filaments were negatively stained with 1% uranium acetate on Ni-400 meshes. A part of the negatively stained preparation was rotary shadowed with platinum, using a BAF060 freeze-etch machine (BAL-TEC AG, Liechtenstein).
Inhibition of actomyosin Mg2+-ATPase activity by TWD2-S
To examine the function of the D2 phosphorylation site in twitchin, a recombinant twitchin fragment (TWD2-S) was prepared that consists of the linker region including the D2 phosphorylation site (S4316) and neighboring Ig motifs (Fig. 1A). Variants of this construct were prepared in which the D2 serine was replaced by alanine (TWD2-A) or aspartic acid (TWD2-D) to represent unphosphorylated and phosphorylated D2 peptides, respectively. The constructs were subjected to phosphorylation assays with PKA to confirm these substitutions. The mutants TWD2-A and TWD2-D were not phosphorylated, as predicted. In addition these data show that no other phosphorylation sites are present in the D2 peptide (Fig. 1B).
ABRM actomyosin Mg2+-ATPase activity in the presence of TWD2-A and Ca2+ was determined to be 40% of the original activity (i.e. in the absence of twitchin peptides) whereas TWD2-D had no effect on activity (Fig. 2A). The molar ratio of the TWD2 peptide to myosin heads in this and subsequent assays was 12:1. In the absence of Ca2+, where molluscan myosin is inactivated, TWD2-A also tended to inhibit Mg2+-ATPase activity, but this effect was not significantly different from that observed with TWD2-D (Fig. 2B). However, these effects were difficult to establish unequivocally since the Mg2+-ATPase activity was low (Table 1). The Mg2+-ATPase activity of chicken skeletal muscle actomyosin (isolated as the complex) in the presence of Ca2+ was decreased to 55% of the original activity, whereas thiophosphorylated TWD2-S had no effect (Fig. 2C). Similar results were obtained with chicken skeletal muscle myosin plus actin (isolated individually) as shown in Fig. 2D. The inhibitory effect of the unphosphorylated TWD2-S on chicken actomyosin Mg2+-ATPase activity was the same in the presence of paramyosin (Fig. 2E) and show that paramyosin has no effect on inhibition by the peptide. Data from these assays are summarized in Table 1.
Binding to myosin, actin and paramyosin of twitchin D2 peptides
It has been claimed that intact twitchin binds to both myosin (Yamada et al., 2001) and actin (Shelud'ko et al., 2004). Thus, the binding of actin, myosin and paramyosin to the TWD2 constructs was investigated using a solid-phase binding assay (see Materials and methods). Thiophosphorylated TWD2-S was used as a probe instead of TWD2-D, because the thiophosphorylated TWD2-S mimicked the phosphorylation form more effectively than TWD2-D. For convenience (i.e. higher yield) we prepared chicken fast skeletal actin and scallop striated adductor myosin. Fig. 3A,C,E show typical results in the solid-phase binding assay for unphosphorylated TWD2-S and thiphosphorylated TWD2-S with actin, myosin and paramyosin, respectively. Unphosphorylated TWD2-S bound to each of the proteins, but not to the thiophosphorylated construct. Relative binding abilities of 10 μg ml–1 twitchin D2 peptides are shown in Fig. 3B,D,F. These results clearly indicate that the twitchin D2 site interacts with actin, myosin and paramyosin in a phosphorylation dependent-manner.
Twitchin binding site on actin
The location of the D2-binding site on actin was determined. Chicken fast skeletal muscle was used as source material to obtain a higher yield. Enzymatic digests of chicken actin were separated by reverse-phase high performance liquid chromatography and each fraction was subjected to solid-phase binding assay with unphosphorylated TWD2-S (Fig. 4A,B). Only fraction 16 reacted with unphosphorylated TWD2-S (Fig. 4C) and this was subjected to a second reverse-phase chromatography. Fraction 16 was separated into three peaks and only fraction 16-2 reacted with unphosphorylated TWD2-S. Its sequence was LVCDNGS, located at the N-terminal end of actin, L10–S16. An adjacent sequence in actin, D26–D27, also is required for the binding of actin to the loop 2 region of myosin and is important for ATP-driven motility (Johara et al., 1993) (Fig. 4D). To investigate whether this region of actin (D26–D27) might be involved in binding to the D2 peptide a synthetic peptide, A21–P29, was prepared and shown to bind to the unphosphorylated TWD2-S but not the thiophosphorylated form (data not shown). This synthetic peptide competed with actin for binding to unphosphorylated TWD2-S (Fig. 4E). The cumulative evidence indicates that L10–P29, exposed on the surface of actin, is involved in interaction with twitchin (Fig. 4F).
Formation of a complex of myosin, actin and TWD2-S
An essential feature of the catch mechanism is the maintenance of tension at low intracellular [Ca2+] where myosin is not active. Do myosin and actin maintain a connection with twitchin during catch? To address this question, myosin and actin were mixed with unphosphorylated TWD2-S under low [Ca2+] and high [ATP] to mimic catch conditions. Due to the requirements for relatively large amounts of myosin and F-actin, scallop striated adductor and chicken fast skeletal muscle were used as starting materials, respectively. The binding of myosin, actin and unphosphorylated TWD2-S was assayed under conditions mimicking catch using low-speed sedimentation at low ionic strength where myosin is insoluble and actin soluble (Fig. 5A). Initially, an excess of unphosphorylated TWD2-S or thiophosphorylated TWD2-S was used (molar ratio for twitchin construct:myosin of 50:9). Thiophosphorylated TWD2-S had no effect on the distribution of actin in the supernatant fraction (Fig. 5A). Cosedimentation of each of the three components was detected only when unphosphorylated TWD2-S was used. Subsequently, it was found that lower ratios of unphosphorylated TWD2-S:myosin also promoted cosedimentation. Even at a ratio of 1:150 (unphosphorylated TWD2-S:myosin) a trimeric complex formation was detected (Fig. 5B). It is proposed that the in vivo molar ratio of twitchin:myosin in catch muscle is about 1:15 (Siegman et al., 1997). These results suggest that under catch conditions the unphosphorylated D2 peptide mediates interaction between myosin and F-actin.
Twitchin localization on myofilaments
We raised an antibody against the twitchin D2 peptide using the recombinant protein (see Materials and methods). The antibody reacted to twitchin, but not to the other major components of the thick filaments, i.e. myosin heavy chain, myorod and paramyosin, as revealed by immunoblotting (Fig. 6A). The antibody reacted with both the dephosphorylated and phosphorylated D2 peptide, but the sensitivity with the dephosphorylated peptide was 10–20-fold higher (Fig. 6B,C).
Transmission electron microscopy was employed to localize twitchin in ultrathin sections of ABRM using the anti-D2 peptide antibody. Different experimental conditions with ABRM were adopted to simulate active contraction, catch state and relaxed state. The abundance of gold particles was highest for ABRM in the catch state (Fig. 7C,D) followed by a decreased distribution in the contracted state (Fig. 7A,B) and a further reduction for the relaxed state (Fig. 7E,F). These data, summarized in Table 2, suggest that twitchin in the catch state is unphosphorylated. Gold particles, which were observed only around thick filaments and not thin filaments, appeared marginally separated from the thick filament surface in active and catch states (Fig. 7B,D), while gold particles in the relaxed state were closer to the thick filaments (Fig. 7F). This slight shift of gold particles indicates that the location of the twitchin D2 site may move at different stages of muscle contraction. No gold particles were observed when ABRM was treated in the absence of the antibody (Fig. 7G,H).
To investigate the arrangement of twitchin on the ABRM thick filaments (isolated from ABRM in the catch state) the distances between closest neighboring two gold particles (recognizing the twitchin D2 site) were measured. These were measured by modifying a measurement position perpendicular to the thick filament axis. A total of 223 measurements were made. Since larger distances are considered to be less reliable for statistical analysis, distances between two gold particles of not more than 400 nm (N=155) were subjected to frequency distribution analysis (Hasselblad, 1966) and separated into a mixture of multiple normal distributions (Fig. 8A). The distribution showed a periodicity of 36.25 nm and the estimated slope of the regression line (36.29) through the origin (Fig. 8B) was not significantly different from the observed period (F-test, P=0.91). This interval of 36.25 nm on the thick filaments is half of the 72.5 nm periodicity of paramyosin filaments (Squire, 1981).
Furthermore, electron microscopy of isolated ABRM thick filaments reacted with the anti-D2 peptide antibody demonstrated that gold particles conjugated with the secondary antibody were located periodically on the surface of the thick filaments (Fig. 9A), confirming the above observations with ultrathin sections. In fact the homogenate of ABRM prepared in the relaxed state contained both thick and thin filaments, but only thick filaments were selected and subjected to analysis. Gold particles attached to the secondary antibody also were associated with granular structures having an average diameter of 20.2 nm, i.e. about the diameter of the myosin head (Fig. 9B), suggesting an interaction between the D2 peptide and the myosin head. Also noted were ultrathin structures of over 50 nm in length and 3.8 nm diameter that extended along the axis of the thick filaments (Fig. 9C). These dimensions suggest, tentatively, that the 50 nm structures may be part of twitchin filaments, extending along the surface of the thick filament. Resolution was not adequate to obtain more accurate estimates of the filament length. ABRM twitchin as observed by rotary shadowing is an elongated molecule approximately 225 nm in length with a spherical head (Fig. 9D), as reported elsewhere (Vibert et al., 1993). The antibody against the kinase domain (Funabara et al., 2001a) reacted to this head region (Fig. 9D). [For these experiments the antibody to the kinase domain was used since its titer for twitchin was higher than the D2 peptide antibody (data not shown).] Models of the parallel array of twitchin molecules (red) are superimposed on the Bear-Selby net pattern (Bear and Selby, 1956) and shown relative to the myosin head distribution (blue) (Fig. 9E). The D2 site is located adjacent to the kinase domain at its C-terminal edge (Fig. 1A) and thus may be positioned close to the myosin head.
In the present study we have shown that the dephosphorylated D2 peptide (consisting of the D2 phosphorylation site, S4316, flanked by two Ig motifs) binds to both F-actin and myosin, but the phosphorylated form does not bind. The binding site on actin was located within the sequence L10–P29 and this region also binds to loop 2 of the myosin head. This interaction between myosin and actin is essential to initiate cross-bridge cycling (Johara et al., 1993). Thus it is expected that binding of either myosin or twitchin to this N-terminal region of actin would be competitive. However, this competition would occur only with unphosphorylated D2 peptide. The binding site for the D2 peptide on myosin remains to be determined, but the electron microscopy data suggests an interaction with the myosin head. As emphasized, the D2 peptide interaction with myosin is phosphorylation-dependent, and it is not known if other phosphorylation-independent links are formed between intact twitchin and the contractile apparatus. The finding that twitchin remains attached to thick filaments isolated under various conditions suggest that there are other twitchin–thick filament interactions (with myosin and/or paramyosin). We propose that a critical feature of the catch mechanism is the phosphorylation-dependent interaction of the twitchin D2 region with both F-actin and myosin. This complex does not involve cross-bridge–actin interactions. The locations of the actin-binding and the myosin-binding sites on the D2 peptide are not known. It is assumed that distinct sites are involved (possibly in the two flanking Ig motifs) and that both are regulated by phosphorylation of D2. It is not necessary that direct binding to the D2 site (S4316) is involved, but this could occur for one of the sites. In addition to twitchin there are several muscle proteins that contain Ig motifs [e.g. titin, myosin light chain kinase (MLCK), telokin and the myosin-binding proteins (MyBPs)] and the ability of certain Ig motifs to bind to myosin has been documented. For example, in cardiac MyBP-C the interaction with myosin was mapped to the C-terminal Ig motif (Alyonycheva et al., 1997). Subsequently, it was found that the unphosphorylated MyBP-C motif bound to the S2 region of myosin and influenced cardiac mechanics. Interestingly, phosphorylation of the MyBP-C motif by PKA abolished these effects by eliminating binding to myosin (Kunst et al., 2000). In cardiac muscle the stoichiometry of MyBP-C to myosin heads is about 1:8 (Kunst et al., 2000). The binding of Ig motifs to actin is not well documented, although the N-terminal two Ig motifs of long MLCK have been implicated in actin binding (Yang et al., 2006).
These results are consistent with those of other investigators (see Introduction) and, for example, may explain the myofilament interconnections proposed by Andruchova et al. (Andruchova et al., 2005). The competition between the unphosphorylated D2 peptide and loop 2 of myosin for the binding site on actin also may be pertinent to the results of Butler et al. regarding the low- and high-force states (Butler et al., 2006). A higher affinity of myosin for actin in the high-force state would effectively displace the twitchin–actin interaction. This is consistent with the observation (Butler et al., 2006) that activation of the catch muscle by Ca2+ increases the proportion of myosin in the high-force state and decreases interaction of twitchin with actin. Transition to the low-force state, by decreasing Ca2+ levels, would favor the twitchin–actin interaction. It should be emphasized that a direct role for Ca2+ in binding of the D2 peptide to either actin or myosin is not suggested and there is no evidence from our data to implicate a Ca2+ sensitive step, other than the regulation of actin-activated ATPase by Ca2+ binding to myosin. Dephosphorylation of twitchin by calcineurin (Castellani and Cohen, 1992; Yamada et al., 2004) would be coincident with initiation of contraction since both events are Ca2+ dependent, and thus for most of the contractile phase twitchin is dephosphorylated and theoretically available to compete with myosin for actin. In practice, twitchin does not inhibit the in vivo contraction of ABRM fibers in the presence of Ca2+ (Siegman et al., 1998). A competition between twitchin and myosin might explain the in vitro inhibition of Mg2+ ATPase activity of actomyosin by relatively high concentrations of TWD2-S.
Results from the localization of twitchin on the thick filament indicate that the twitchin molecules are distributed on the surface of the filament at a periodicity of 36.25 nm, half of the 72.5 nm periodicity of paramyosin filaments (Squire, 1981). It is suggested that the D2 site at the C-terminal end of twitchin aligns with myosin heads, i.e. at half of the 72.5 nm axial periodicity (Cohen, 1982). In the model proposed in Fig. 9E the twitchin molecule could extend through about three of the 72.5 nm repeats or nodes, if arranged parallel to the filament axis. The myosin rod extends axially through two nodes. Thus there is a possibility of other interactions between twitchin and the myosin rod. The molar ratio of twitchin to myosin is about 1:15 in ABRM catch muscle (Siegman et al., 1997). Therefore, even if all of the twitchin molecules interact with myosin, the number of trimeric complexes is small compared with total myosin. It is not known if there are cooperative effects on neighboring myosin molecules induced by the binding of twitchin to myosin. The ultrathin filaments observed above on the surface of the thick filaments are thought to be twitchin and these reflect the underlying Bear-Selby net (Bear and Selby, 1956) (see Fig. 9C).
There are several aspects of the catch mechanism to be resolved. In an earlier report (Funabara et al., 2003) it was stated that phosphorylation of both the D1 and D2 sites is required for relaxation from the catch state. The D1 site is unusual in that relatively high levels of phosphorylation (40–50%) are found while catch is maintained. The D2 site is more sensitive to effects of phosphorylation; low levels of phosphorylation are found during catch and high levels of phosphorylation accompany the release from catch (Funabara et al., 2003). The more marked phosphorylation dependence of the D2 site prompted this present study, but the role of the D1 site is not understood and should be resolved by further studies. Another area for future study is to identify the phosphorylation-dependent binding site on myosin for the D2 peptide and also to determine if other interactions between native twitchin and myosin occur. If present, these could anchor the twitchin molecule to the surface of the thick filament. Based on the in vitro results of Yamada et al. (Yamada et al., 2001) it is assumed that paramyosin is not an essential component of the catch mechanism, but whether it has any influence on catch under in situ conditions remains to be determined. The phosphorylation-dependent binding of the D2 peptide to paramyosin is shown above. It is known that in fibers of molluscan smooth muscle the catch state is sensitive to pH (Hopflinger et al., 2006) and is reduced on moderate alkalinization to pH 7.2–7.7, i.e. at those pH values used for the binding experiments described above. It is important to determine if any of the interactions between isolated proteins show a marked pH dependence and this can be tested experimentally. On the other hand, the sensitivity to pH may be associated only with the intact contractile system. Recently, it was found that striated muscles from oyster and scallop contain twitchin and this regulates interaction between thin and thick filaments at low [Ca2+] (Tsutsui et al., 2007), as it does in molluscan catch muscle. These results suggest a similar role for twitchin in striated and smooth molluscan muscle and the molecular scheme outlined above may be applicable to both muscle types.
In summary, our results provide novel data on the molecular interactions involved in catch. In Fig. 10A the different phases of the contractile cycle in catch muscle are shown, with associated changes in [Ca2+], tension and twitchin phosphorylation. Starting with relaxed muscle (Fig. 10B), twitchin is phosphorylated (by PKA) and the D2 peptide does not interact with either actin or myosin. It is assumed that in the relaxed state there are no interfilament connections. Stimulation by acetylcholine increases [Ca2+], which promotes contraction (by binding to and activating myosin) and activates calcineurin. Twitchin is dephosphorylated but does not bind to actin (although theoretically this interaction is still possible) because of competition with the increased population of high-force states, i.e. the cross-bridge–actin interaction predominates. The onset of catch follows a reduction in [Ca2+] to close to resting levels and inactivation of myosin. The low-force state (the detached cross-bridge being part of the low-force state) allows the D2 twitchin region to bind to F-actin and myosin and form a mechanical force-bearing complex. In this model there is no direct effect of Ca2+ on the binding of the unphosphorylated D2 peptide to either myosin or actin and our hypothesis is that the obligatory role for a reduction in Ca2+ to promote the catch state reflects the inactivation of myosin and an increase in low-force state. This complex is proposed to represent at least part of the mechanical connection (catch bridge) between the myofilaments. Serotonin release causes an increase in [cAMP] and activation of PKA. One of the PKA sites on twitchin is the D2 site and its phosphorylation eliminates binding of the D2 peptide to actin and myosin. The catch connection is lost and the muscles enters the relaxed phase.
We thank Dr M. J. Siegman and Dr T. M. Butler for their advice and Dr Takashi Yamakawa for help in statistical analysis. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to D.F.) and Japan-US Cooperative Science Program from Japan Society for the Promotion of Science (to S.W.) and NIH grant HL-23615 (to D.J.H.).
↵* These authors contributed equally to this work
- © The Company of Biologists Limited 2007


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In the first of a series of profiles of designers taking part in the Local Designer Showcase, Bernews today spotlights Satara Caisey. The Local Designer Showcase is set to be held on July 10, 2014.
When asked about her reasons for participating in the Local Designer Showcase, Ms. Caisey said, "I saw this as an opportunity to introduce myself to Bermuda as a designer. As a recent Fashion Design Graduate, this is a great way to network when first coming back from university."
When asked what she hoped to get out of the experience, Ms. Caisey said, "I hope to meet like-minded people. I believe this experience will also give me an opportunity to see the Bermuda market from the perspective of the designer. I currently work as an Administrative Assistant Buyer and that has afforded me the opportunity to have insight into the Bermuda consumer."
When asked what progress had been made so far on the completion of her line, Ms. Caisey said, "My line is almost complete. From my graduate collection, I learned that you need a time line and have to stick to it, because you never know what will change.
"For instance, this was the first time I have had to look for sponsorship, and I learned early on it wasn't going to be easy. So you have to do as much as you can with what you have. If and when sponsorship comes, use it wisely. For example, I initially wanted to print onto fabric, which is a costly process. You have to be realistic with your budget, you can't bank on getting sponsorship."
When asked what her favorite piece in her new line was, Ms. Caisey said, "I wouldn't say I have a favorite piece. They have all evolved so much from concept to creation. Sometimes you design something on paper and you find a fabric that you love but the fabric doesn't work well with the design.
"So you have to choose; you choose between the design and the fabric. This is why I try to choose fabric first. However, when you walk into a fabric store you never know what will catch your eye on your second or third visit. Things change."
When asked what her experience with her mentor was like, Ms. Caisey said, "It has been an amazing opportunity to meet someone in the industry. It is very rare that you get to meet someone, nevermind have direct access to a person, with such experience. Also the check in's helped to stay on target."
When asked how she knew that design was her passion, Ms. Caisey said, "The arts have always been a passion of mine. In high school, I took Art and Design, Textiles, and Design Technology. However, when I graduated, while I was passionate about the arts and fashion, I took what I suppose I thought was the safe and smart route. I decided to study law. While I enjoyed law, as time went on I knew I didn't feel fulfilled.
"I completed my law degree and graduated in 2009, but after graduating I took a year off. During this year I took short courses in Draping, Pattern Cutting, Styling and Fashion Design, and Marketing at Central Saint Martins. I knew I had to be sure exactly what in fashion I wanted to do before starting another degree. I decided on a Fashion Design BA at Middlesex University and graduated last year."
When asked what the future holds for her as a designer, Ms. Caisey said, "While I don't know what the future holds, I have goals that I aim to achieve. Two of them are being realized now. One was to be a part of this fashion show. I went to the show last year and less than half way through I knew I had to be a part of this experience.
"And the second was to find a job in fashion in Bermuda. I say in Bermuda because that was a big part of the goal. I hope to see my garments in stores as well as eventually own a store of my own. This will house not only my designs but the designs of fellow Bermudians, whether they live here or abroad."
When asked how she would describe her sense of style, Ms. Caisey said, "My style would probably be understated chic. I am actually trying to introduce colour into my wardrobe now. I tend to have blacks and greys in the winter., and beige, whites, and natural colours in the summer."
The City of Hamilton will play host to the second edition of the Local Designer Showcase on Thursday, July 10, 2014 at Pier Six [upper level], with eight local designers to be showcased for their talent and unique work. Each designer has been peered with a mentor from within the fashion community to assist them with the development of a fashion line that will be showcased at the event.
From: Bernews.com


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Foreclosure Rates, How Sweet the Sound, that Saved a Wretch Like Me
I'm on the record as a skeptic of predictions that we'll start bowling together again once we've been downsized out of our McMansions and SUVs. But this article suggests that suburban neighborhoods are actually responding like the optimists hope. Aside from the anecdotal evidence, and the gauzy hopes of urbanists and sociologists, there appears to be a genuine uptick in local organization. Suburbia, per Joel Kotkin, could have the last laugh yet.
But get this:
"It's more caring. . . . It's like we're a team," said Rob Burnett, 44, of Fredericksburg. Residents in his subdivision have planted trees, donated items to a neighbor with five children who lost his job, and formed a study group on debt-free living. "Before, everybody was showing off what they had. Now it's like, 'What can I cut back?' and 'How are you doing things differently?' Before, the guy who has the biggest Hummer on the street was the biggest guy in the world. That's gone."
I read that and my first thought was to bang my head on the desk in embarrassment. "Unemployment rates and underwater mortgages have turned you and your neighbors from an enclave of self-absorbed, positionally obsessed, Hummer-worshiping Patio Men into a real community overnight? How sweet." Did it really take a dire macroeconomic backdrop to inspire them to help an unemployed neighbor with five kids? The cartoonish self-congratulation in his conversion story makes me distrust both Burnett's account of yesterday's groveling materialism and his praise for today's virtuous austerity.
But you go to war with the army you have, and if Americans are going to pull through a tough recession and discover the virtues of mutual support and community engagement, it will probably not be through a wholesale reorganization of our priorities — it will happen in a way that flatters and accommodates our peculiar (and peculiarly American) flaws and fantasies. Rather than deep moral and spiritual renewal leading to civic health, what if it's our national solipsism and susceptibility to suggestion that pull us together, and pull us through? What if, rather than being stuck with virtue, we discover that, after a few initially painful changes in lifestyle, we can buy spray-on virtue in a can? If enough Americans decide that the TV show of their lives should feature them acting like engaged, conscientious citizens, might that not be just as good as a more "authentic" conversion?
It might be exasperating to live among neighbors who are acting out a self-conscious "sense of" community, but that may be the precise way our better natures come to light these days. And the available alternatives could be a hell of a lot worse.


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Love is the theme.
Love is the theme.
When Lana Del Rey isn't gathering the troops to cast a spell on Donald Trump, she's perfecting her craft. During a creative session with English musician The Rascals and The Last Shadow Puppets' Miles Kane in L.A., the songstress took a moment to call into BBC Radio 2's Jo Whiley and explained how the world's volatile social and political environment inspired her forthcoming record.
"I started out thinking that the whole record was gonna have a sort of a '50s-'60s feeling, kind of some kind of Shangri-Las, early Joan Baez influences," she said. "But I don't know, as the climate kept on getting more heated politically, I found lyrically everything was just directed towards that. So because of that, the sound just got really updated, and I felt like it was more wanting to talk to the younger side of the audience that I have. I guess it's a little more socially aware. It's kind of a global feeling."
Listen here:
Thoughts? Let us know in Exhale.
And while you're at it… leave a comment and follow us on Facebook, Twitter & Instagram and subscribe to our YouTube channel.


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S6796 - Industrial Land for Sale - SGN
Gerehu, Port Moresby, NCD, Papua New Guinea
Located in the Gerehu Industrial hub is this vacant industrial land immediately available for sale. Land size is 0.240 ha and is fully serviced, gated and surrounded by prominent business houses. An ideal opportunity for an investor involved in industrial related business activities similarly in conjunction with the new proposed Port at Motukea. The vendor is keen to sell and genuine offers will be considered. Please contact the undersigned sales agent to schedule an inspection.
Property Information
- Property Type: Land
- Property ID: 1623
- Original ID: 2344180
- Price: K2,500,000
- Floors: 1
Property Features
- Close to Transport
- Quiet Location
- Safe
Location Profile
Port Moresby
Port Moresby Overview Port Moresby is the capital of Papua New Guinea (PNG) and is the largest city in the Pacific Islands outside of Australia and New Zealand. It is located on the south-eastern coast of the Papuan peninsula on the shores of the Gulf of Papua on the island of New Guin...View location profile
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30 years
Repayment K500 / week
Century 21 Siule Real Estate was established in 1973. By being SMARTER BOLDER FASTER, we have become the most successful and most professional realtor in the unique environment that is PNG real est...


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A new 30-second public service announcement encouraging people who are building new homes to ask for home fire sprinklers has been making the rounds on HGTV. The spot, created by the non-profit Home Fire Sprinkler Coalition, was featured during popular programs like "Property Brothers", "Curb Appeal", "Love It or List It", and "House Hunters International", and was expected to reach some 32 million viewers with its important message.
"Thinking of building a new home? Now is the time to think about protecting it with fire sprinklers. Nine out of 10 fire deaths happen where you least expect them, at home. And those at greatest risk are children, older adults, and people with disabilities. Despite what you see in the movies, the entire system does not go off; only the sprinkler closest to the fire activates and stops the fire from becoming deadly. If you are building a new home ask for fire sprinklers to protect your family and your home."
If you're a homeowner or someone who's looking to build a new home, the Home Fire Sprinkler Coalition has free online materials to help you understand why fire sprinklers are needed, how they work, and how they are installed.


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- Description
* Deep dark red finish with replicated mahagony grain.
* Richly finished beads, carvings and rope details throughout the collection.
* Enormous poster bed with storage footbaord option.
* Framed top drawer with glided bead detail.
* Glossy faux marble tops.
* Side roller Glides for smooth operating drawers.
* Fluted bun style feet support the cases.
* Detailed antique gold color hardware.
* Abundant shape and detail across the dresser inlcluding a stepped out middle section.Dimensions and Weights:
Item Dimensions Weight Gabriel King Poster Headboard 82"W X 5"D X 71"H 125 lbs Gabriel King Poster Footboard 82"W X 8"D X 34"H 110 lbs Gabriel King Poster Rails 79"W X 84"D X 18"H 40 lbs
- Additional Information
Additional Information
Color No Ship Time 5-10 Business Days Items Included King Poster Bed Design By No SKU # B347-68-66-99 Manufacturer Ashley
- Reviews
- Coordinating Items
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Archaeology & Spirituality
Many believe the West Penwith peninsula is a spiritual place. Its remoteness, its stark beauty, its presence of nature, and the many archaeological treasures that are present in the landscape, connect us with this place and with the people who have previously lived and worked this land.
A patchwork of small fields and granite Cornish hedges reflect the prehistoric field patterns bordering the Penwith Moors and provide a unique insight into an agricultural landscape which is between two and three thousand years old.
There is a wealth of archaeology to be explored on foot: the outcrops along the coast with remnants of cliff castles, the raised beach of Porth Nanven (Cot Valley – it shows climate change in geological times), Neolithic and bronze age cairns, tumuli and barrows (Ballowal Barrow), Men-an-tol, Carn Euny, Chun Castle & Quoit, stone circles such as at Tregeseal, standing stones, fogous (pronounced "foo-goos"), celtic crosses, and holy wells.
There are numerous granite churches that are so characteristic for the Tin Coast and surrounding area, for example in St Just, Pendeen, Morvah, Zennor, Paul, St Buryan, and Madron, where people have worshipped to this day.
The St Just Miner's Chapel is one of the best examples of Wesleyan Methodist Heritage in Cornwall. Its graveyard reflects the life of the characters who inhabited this mining town, and are part of people's family history. In its heyday, the 'miner's cathedral' would have been filled to the brim with people and song, but recently the congregation has been dwindling. The local community is keen to 're-purpose' the building, so it once again becomes the centre of the community.


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1111 Nw 85th St, Seattle, WA 98117
Toll Free: 888-782-4213 Local: 206-782-4213 Fax: 206-784-0535Email Us
- 1111 Nw 85th St, Seattle, WA 98117
Wreath of white lilies, roses and flowers.
All items featured on this site represent the types of arrangements we offer and may vary depending upon availability in certain regions. Substitution Policy


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Several thousand patients a year in the UK have Arthroscopic procedures – keyhole surgery, either on the NHS or privately. One of its indications is to reduce knee pain caused by Osteoarthritic changes in the knee by tidying up damaged cartilage and ligaments or removing loose debris and floating bits of cartilage from inside the joint. The operation involves making two small cuts on either side of the front of the knee. One is for a telescope – Arthroscope, enabling the surgeon to see directly inside the area, and the other is for instruments to carry out the surgery.
Researchers in America compared real surgery in 180 patients with a placebo group of patients in which the patients were given an anaesthetic to make them fall sleep and a local anaesthetic to make the skin incisions. In the placebo group the surgeons did not go into the knee joint.
All the patients reported reduced pain and better knee function, and there was no real difference in outcome between those who had real surgery and those who got the dummy or placebo procedure at the two year follow up.
For the younger non arthritic patient with ligament or cartilage damage there is still a strong case for keyhole surgery, but this new research may start to alter current thinking of using keyhole surgery as intermediate line of attack for the Osteoarthritic joint to one of a just recommending a partial or full knee replacement.[ngfb buttons="facebook, gplus, twitter, linkedin, pinterest, stumbleupon, tumblr"]


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Bradley Smoked and Grilled Lamb Neck Fillet with Mediterranean Vegetables
Sometimes just a subtle hint of smokiness can transform a dish. Lamb can easily be spoiled by too much smoke, and needs to be served pink to retain its juiciness so the solution is to lightly smoke it before briefly grilling it over intense, direct heat. This is an ideal recipe for the Countertop Bradley Smoker, or the standard versions. writes Sandra Tate … Lamb rub For this recipe you need to prepare a fragrant rub for the lamb, this amount will be sufficient to coat 2 whole lamb neck fillets. Ingredients: 2 whole lamb neck fillets 10g sea salt flakes (Maldon or similar) 5g toasted fennel seeds 1 tbsp finely chopped rosemary leaves 1 tsp honey 1 tbsp olive oil 1 large clove garlic, peeled & crushed finely grated zest of half a lemon Place all the ingredients in a mortar and grind together with a pestle until you have a paste. Divide the rub between the two neck fillets and coat evenly all over. Cover lightly with cling...Read More


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This is the fourth of five qualifying Smartypants quizzes this week, all leading up to one big fat Large Marge Smartypants Quiz on Friday night.
Two winners of this quiz will win spots in tomorrow night's quiz.
Quiz time will end at 11:30 pm Pacific Time Thursday night. (Answers will be hidden until tomorrow morning. You will be able to see your answers, but no one else will!)
Winners and answers will be announced at 8:00 Pacific Time Friday morning; Quiz #5 will be posted at 9:00 am Pacific Time Friday morning.
Good luck!
1. What communicable disease is also known as pertussis?
2. What is the name of Valerie Bertinelli and Eddie Van Halen's son?
3. Which U.S. president was the father of Maureen, Christine, Michael, Patti, and Ron?
4. What breed of dog was Rin Tin Tin?
5. Who is this woman?
6. Identify these creatures:
7. Who is the lead vocalist of the rock band Rush?


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Home » Quilters supplies
Notions, tools, supplies, thread & quilting accessories for the quilting enthusiast. Including rulers, cutting mats, rotary cutters, scissors, seam rippers, pins, fabrics and more.
+ Special Discounts to Schools, Theaters, Dance Groups, Weddings, Caterers, etc.
Fabrics & MoreBringing Your Ideas to Life
Keeping our loyal customers happy for 50 plus years with excellent customer service and selection!
Copyright @ 2017 - Yardage Town - Fabrics & More Bringing Your Ideas to Life!


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Our resident involvement strategy
In order to achieve our strategic plan, we must work to support strong resident groups and engagement. These empower residents to shape the services which work for them. We must also support community initiatives. This includes groups which help residents to create lasting, positive social change in their neighbourhoods.
Why it is important for residents to get involved
You know best how to make things better, because you receive the service that we provide. You live in the communities and areas we serve. No matter how big or how small you can make a difference. You can get involved by helping improve the services you receive from us. Or helping change and improve the area you live in.
By giving some of your time we can find out what is good or bad about what we do. What works and what doesn't. Everyone has an opinion and we want to listen to yours so we can improve. It's not only about listening but also ensuring what we say we'll do, we actually do.
Support and help
We'll do everything we can to give you the support you need to feel comfortable about getting involved. Don't worry if you can't get to a meeting or can't afford bus fare or childcare. We can always help arrange transport. We will cover any reasonable expenses as long as you provide a receipt.
Ways to get involved
Below are examples of opportunities available to residents at the moment. Also, roughly how much time each activity requires. We appreciate that not every option will be suitable for everybody. Hopefully there should be something for all. If you want to be involved in a way that isn't included in the list then let us know. We can see what we can do to accommodate you.
Do you have an idea for your local area? Want to get more involved with your community? Or be part of a group looking to improve services for residents? Please come to the Garth and talk to the Resident Involvement Officer or Community Development Officer, or contact us.
Board member
The Board ultimately is responsible for everything that we do. Board members ensure we act in accordance with the law. And also, the regulatory standards that are set for social landlords.
Time commitment:
Full Board members are likely to commit at least 146 hours per year to carry out their role effectively. This could be as much as 200 hours in a very busy year.
Tenants & Resident Scrutiny Panel
Resident Scrutiny Panel (TRSP) is a group of residents from rented, shared ownership and fully owned homes. They live in different communities. The TRSP work together to review different service areas. They make recommendations for change and improvements based on the evidence they find. They present their findings to the Board.
Time Commitment:
Two to three hours a month for monthly meetings. Additional time will be required whilst completing a project, training or reading paperwork.
Focus Groups
We regularly get together groups of residents who look at specific service areas. These are usually residents who have received the service, or the service is relevant to them. In the past, focus groups have looked at our strategic plan, modernisation, gas checks policy and VOIDS. A good way of getting involved if you don't have time to join a group on a regular basis.
Time Commitment:
Each focus group lasts approximately one to three hours.
News panel
Our resident magazine is sent to all residents. It is produced three times a year. The news panel consist of both staff and residents. The panel meets before each issue to discuss content. This includes information for, and stories about, our residents and the latest news in the world of housing. The panel then comments on it before it goes to print.
Time Commitment:
One lunchtime meeting per issue, each approximately two hours. Additional time to look at and edit the final version.
Local resident groups
There are a number of existing resident groups. They are made up of tenants and home owners who meet regularly to talk about local issues. They can make, and have made a huge difference to the communities they are based in.
Time Commitment:
One meeting once a month, for approximately one to two hours.
To find out if there is a resident group in your area, or to set up a group, contact us.
Residents Voice
The Residents Voice group represents resident groups and committees across all our communities. It looks at decisions about improving housing and neighbourhood services. As part of the group you will be representing your community, scheme or area. The meetings are to share news with other residents and feedback to us about our services.
Time Commitment:
Three meetings a year, approximately two to three hours. Can be at various locations.
Resident Advisor
These are volunteers who work closely with staff. They help to monitor and develop our services in a particular area of operations. Resident Advisors receive appropriate training to prepare for their role. This is arranged and paid for by us. Typically Resident Advisors assess an agreed aspect of a service. They read documents and carry out interviews, surveys, phone calls and personal visits. They then make recommendations on how the service can be developed. They identify both the positive and negative aspects. They can also summarise the comments of other residents.
Time Commitment:
Average of one or two days per month.
Community events and groups
We encourage and support wider community involvement. This includes social events, interest groups, environmental projects and events for unrepresented groups and individuals. This includes children and families.
You can also set up or join a 'walkabout' in your community. This is to point out areas of concern about cleanliness and maintenance of your area.
Resident surveys
Resident satisfaction surveys are undertaken for some of our services. Please take the time to complete and return these to us.


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"Nobody seemed to be as good in moving as with Metropolitan Movers. I had a couple of experience with other companies before and they were both nightmares. When I started hiring these guys, I was totally shocked at how convenient relocation can be if you are with the right team. Will definitely stick with them. I suggest you should too!"
"Just had another pleasurable move with this company. Nothing but praises for their service and professionalis. Well done, as always. Regular client here. "
"The move these guys did for me was absolutely incredible. The arrived on time, they were very experienced and organized and so my move is completed in a blink of an eye! And I thought, moving was stressful, NOT with Metropolitan Movers."


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It's hard to get out of Alphabet City these days, now that it's the hipster eating-and-drinking mecca. Unlucky for us, the ABCs are ridden with a glut of generic lounges. Don't these people have any imagination? Do we really need another no-dancing zone that teases us with pounding bass? No, Liquid City denizens, we do not. So thank your stars that some of the recent additions have that certain je ne sais quoi. Many are run by the city's latest wave of hardworking immigrants and are closer to Avenue C, where the rents are a little cheaper and the draw (as well as the drinks) needs to be a little stronger.
Smoke your Gitanes while looking bored and breathless at BELMONDO (98 Avenue B, 358-1166). Simple, open, and airy, this new provincial bistro is just as cool, but not nearly as pretentious as its über-sexy namesake, French acteur Jean-Paul Belmondo. Go incognito with a Godard ($7)—a passionate mix of Absolut Kurant, Chambord, and pineapple juice—or a refreshing, minty mojito ($7). The cocktails taste best at dusk, when the soft glow of flickering fireflies, yellow bug lights, and aquamarine tiling offsets the midnight blue sky on view from the lunch-counter-style bar. Friendly service and reasonably priced café standards like croque monsieurs ($9) and steak frites ($15) make you go ooh-la-la.
Rock the casbah at RICO CAFÉ (153 Avenue C, 505-5757), where the Egyptian shishas ($7), or hookahs, get packed with banana tobacco. The eclectic storefront is a hodgepodge of exotic artifacts, mismatched chairs, and exposed brick. But what sets this place apart is its hospitality: Brightly colored sheaths fit for Tutankhamen were slipped over our heads, and when the funky Arabic sounds of al-jil got us up and writhing, our host, Mike, Prince of Egypt, joined in and taught us some moves. Alcohol's a Muslim no-no, but, hey, this is Alphabet City, so Heinekens and Coronas ($4) are on offer.
Gothic dungeon SHAMPU (9 Avenue A, 646-602-2590)—with its dripping candles, crimson lights, and brick and wood-paneled interior—seems more a setting for s/m than pre-club cocktails. But the plushy couches and disco ball say otherwise. A recent Friday found an efficient yet unremarkable bar staff pouring the usual screwdrivers while a well-heeled mix of house lovers (Jersey Italians, Queens Koreans, and Uptown Puerto Ricans) chilled out before a big night on the town. Warning: Move on before the clock strikes twelve, when the crowd gets thick and full of testosterone and the DJ'ing starts sounding like Hot 97. Some meatheads gave this critic static when she tried to get up to the bar to settle the tab. Apparently, her pocketbook brushed against one of them. Imagine the damages they sustained! Good citizens of Liquid City: I'm too old for this bullshit!
This article from the Village Voice Archive was posted on July 10, 2001


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Serpent Sweatshirt
Serpent Sweatshirt
Slick Lane 2019 Premium Collection Features: Handcrafted using a premium blend of polyester and cotton (70% polyester, 27% cotton, 3% elastane) which provides the perfect canvas for high-quality sublimation prints. As a result, the artwork will never...
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Slick Lane 2019 Premium Collection
- Handcrafted using a premium blend of polyester and cotton (70% polyester, 27% cotton, 3% elastane) which provides the perfect canvas for high-quality sublimation prints. As a result, the artwork will never crack, peel or fade, even after multiple washes!
- Soft cotton-feel face with brushed fleece fabric inside
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- Regular fitting pullover sweatshirt
- Standard US men's sizing. Ladies, we recommend that you order one size below your usual size. (Please refer to the sizing chart)
Free shipping
7-19 Business Days
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Free shipping
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Free shipping
7-19 Business Days
Please be aware that the shipping times may be delayed due to unforeseen circumstances such as customs processing delays which is unfortunately not under our control.
You will receive tracking information 3 to 8 days from the date of purchase. If you have not received any tracking information after 8 days, please notify us via email (email@example.com).
Please request a refund or an exchange by contacting us via email (email@example.com) within 30 days of the delivery of your product. If the return is a result of our error (you received an incorrect or defective item, etc.) we will also refund the shipping costs of returning the item.
Slick Lane reserves the right to refuse a refund or exchange if the item in consideration is deemed by us to be damaged by the buyer.
Please check out our FAQ page for helpful answers to further questions you may have.


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Our firm
Our firm
For more than 80 years, La Méridionale has been a key figure in Marseille. Today it is one of the oldest ship-owners in the town. Its activities have developed considerably since it was founded. Originally an international freight carrier, it now runs a service to Corsica every day as well as a twice-weekly service to Sardinia for passengers and freight
To find out more about us, consult the different sections.


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Photo Credits:
Brittney Mooney Photography
Our Studio
Providing technically proper body alignment training, and age-appropriate movements in a nurturing, family atmosphere. Lanigan Dance Dynamics programs are for students of all ages who seek personal enrichment through dance training. Our goal is to bring out the best in our students, and help them to reach their full potential. We do this by offering the best in technique and the newest choreography, while cultivating a love for dance in our students. We strive to raise the bar on excellence in dance education while providing a safe environment fostering achievement at the highest levels.
PO Box 222 Lanigan, SK S0K 2M0 | 1-(306)-365-4344 | email@example.com


text: Whether funding, acquiring, managing or disposing of assets, lenders, investors and their advisors can use CoStar to manage risk and analyse opportunities; benchmark and report on the performance of assets; and significantly speed up their CRE research process. CoStar Group provides transparency and drives efficiency in the commercial property sector through real time data and insight. Thousands of property professionals now use CoStar, where 500,000 commercial buildings are tracked for investment and lending opportunities; space availability and deals; tenant movements and lease events.


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Arsenal returned to the top of the Premier League with a nervy victory over Cardiff City on New Year's day, and while Nicklas Bendtner and Theo Walcott found the net for the Gunners, talk afterwards focused on the club's need to recruit additional stars in the January transfer window.
Although some continue to question the need for new faces, Arsene Wenger has admitted he will be "on alert" over the next month for potential arrivals.
"We are like many clubs: certainly on alert if something special turns up and we will not turn it down, but we are not desperate because we have a big depth in our squad," he said.
As such, a number of reports have surfaced linking Arsenal to a whole host of stars, and talkSPORT have handily waded through the mire to bring you the latest news on the club's potential arrivals.
Click the arrow on the right for more.
Arsenal fans, who would you like to see arrive in January? Comment below…