text:
Boost Your Body's Metabolism
At Heat Fitness, our Thermal Pods create the optimal workout environment for your body. Developed in Switzerland and refined in Italy and Argentina, this technology helps you build more strength, tones stubborn "problem areas" and melts inches like never before.
Each pod is heated to 100 degrees Fahrenheit to match peak body temperature. This burns more fat and calories during your workout and boosts your metabolism so you'll continue to slim down even after your session.
Combined with our hands-on personal training and high-rep, low-weight workouts, Heat Fitness' Thermal Pods can give you a leaner, more toned appearance after only a few sessions.


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Trinity House - Procurement Portal
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UK-London: Demolition, site preparation and clearance work.
Preliminary Questions
This notice is for prior information only: Yes
This notice aims at reducing time-limits for receipt of tenders: No
This notice is a call for competition: No
Section I: Contracting Authority
I.1) Name and addresses:
The Corporation of Trinity House
Tower Hill, London, EC3N 4DH, United Kingdom
Tel. +44 1255245112, Email: email@example.com
Contact: Beth Davis
Main Address: www.trinityhouse.co.uk, Address of the buyer profile: http://trinityhouse.g2b.info/
NUTS Code: UKI
Specifications and additional documents: As Above
I.2) Joint procurement:
The contract involves joint procurement: No.
The contract is awarded by a central purchasing body: No .
I.3) Communication
Further information can be obtained from: the above mentioned address
I.4) Type of the contracting authority
Body governed by public law
I.5) Main activity:
Other activity: Maritime Navigation
Section II: Object
II.1) Scope of the procurement - 1
II.1.1) Title: Decommissioning of Royal Sovereign Lighthouse
Reference number: T0389
II.1.2) Common Procurement Vocabulary:
45111000 - Demolition, site preparation and clearance work.
II.1.3) Type of contract: WORKS
II.1.4) Short description:
Trinity House is looking to appoint a contractor to deconstruct, remove and dispose of the Royal Sovereign Lighthouse structure.
II.1.5) Estimated total value:
Value excluding VAT (give figures only) : 8,500,000 - GBP
II.1.6) Information about lots:
This contract is divided into lots: No
Maximum number of lots that may be awarded to one tenderer: Not Provided
The contracting authority reserves the right to award contracts combining the following lots or groups of lots: Not Provided
II.2) Lot Description
II.2.2) Additional CPV code(s):
45111000 - Demolition, site preparation and clearance work.
II.2.3) Place of performance:
Region Codes:
UKJ21 - Brighton and Hove
II.2.4) Description of the procurement:
Trinity House is looking to appoint a contractor to deconstruct, remove and dispose of the Royal Sovereign Lighthouse structure.
II.2.14) Additional information
Not Provided
II.3) Estimated Date of publication
Section IV: Procedure
IV.1) Description
IV.1.8) Information about Government Procurement Agreement (GPA )
The contract is covered by the Government Procurement Agreement (GPA) - NO
Section VI: Complementary Information
VI.3) Additional Information:
For more information about this opportunity, please visit the Delta eSourcing portal at:
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VI.5) Date Of Dispatch Of This Notice: 28/06/2019
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text: President Bill Clinton promoted General Franks to fours stars. President Clinton also appointed Tommy Franks as Commander-in-Chief of the United States Central Command beginning in July 2000. General Franks served in that role through July 2003. In between was 9/11. Tommy Franks led the American and Coalition forces to victory in both Afghanistan and Iraq.… Continue reading American Soldier by General Tommy Franks


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Les Chalets sur le Cap
Establishment No : 136452
Our typical Québec-style cottages, completely built with white pine, combine traditional charm and modern comfort. Located on a high cape, visitors will enjoy the magnificent view on the St. Lawrence River. On site, visitors will appreciate the nature trails in the forest and the relaxed ambiance. Mont-Sainte-Anne, Cap Tourmente wildlife reserve and Canyon Ste-Anne are 5 minutes away. Just 25 minutes from Old Québec City and 20 minutes from Le Massif de Charlevoix.
Units: 7
Some unit(s) available with functional fireplace
All nonsmoking establishment
Room(s) with kitchennette available
Room(s) with scenic view
Online reservation possible
Internet Access
Contact Information
201, route 138, Saint-Joachim, QC, G0A 3X0
Telephone : 418 827-6777


text:
- 沖縄県正社員外資系企業
Lead and manage financial responsibilities at a top American hotel brand in Japan. You will support the expansion of a top international firm in growing their Japanese business and start new projects to develop a new team. You will be working as close partners with the CFO to develop reports to the headquarters in the US.
求人を保存 求人を見る
Support the start of new projects and monitor to develop the company business
Collaborate with top class coworkers in a diverse and multinational environment


text:
Posts Categorized: Medical Device Industry Topics
How to Advance Your Medical Device Career in Southern California
The medical device field is booming. Great leaps forward in medical technologies, combined with an aging population, mean the demand for new and innovative medical devices will continue to climb in the coming years. Whether you're already in the medical device field or wish to make an entry, here's how to advance your career with… Read more »
How to Attract and Hire Passive Candidates in the Medical Device Industry
The medical device industry is booming as new research and new applications of existing devices are uncovered and developed into the means to treat a wide range of pressing medical conditions. Since company need for these medical device professionals is currently outpacing available talent, being able to find great candidates is a must. If job… Read more »
How to Find a Great Job in the Medical Device Industry in SoCal
The medical device industry is booming. Many companies in Southern California work within the medical device industry, taking innovative approaches to healthcare and developing breakthrough treatments to improve quality of life for ill or disabled individuals. If you're looking at the medical device industry for your next career move, here's how to find a great… Read more »
Career Tips to Break Into the Medical Device Industry
The medical device industry is expanding rapidly as advances in science, medicine, and technology open up previously unexplored areas of medical care and treatment. For IT professionals, marketing and sales professionals, and others, work in the medical device industry offers the opportunity to track your career growth alongside the growth of a fascinating and fast-paced… Read more »
Improve Your LinkedIn Profile by Getting Published
A publication credit on your resume improves your standing in the medical device industry. It demonstrates that you've gained the expertise to offer real value to your readers, and it shows that you have the passion and dedication to sit down and put that expertise into writing. If you've focused on honing your skills in… Read more »
Medical Device Tax – Update & Talks of Change
When the Affordable Care Act became law in 2013, it changed the tax code in a number of ways, including the creation of a tax on medical device sales. At the time, the medical device tax was thought to be only a minor part of the bill – but the push to repeal it has… Read more »
How to Increase Retention Rates in the Medical Device Industry
If you've tried to hold on to good medical device talent, you know how challenging it can be. The cost of turnover is higher than in other fields because the knowledge base you lose can take a long time to replace. The complexity of medical devices means it takes longer to achieve expertise. Unfortunately, many… Read more »
How to Recruit, Hire, and Retain Top Talent in the Medical Device Industry
The medical device industry is one of the fastest-growing industries in the United States, and the pressure is on companies large and small to find and keep the best talent available. Here are some "top tips" for top talent: Recruit Know who you need. The more information you can provide your staffing firm about the… Read more »
Three Traits That Promise Success in the Medical Device Industry
The medical device industry is growing strong, leaving many companies scrambling to find qualified candidates for a number of positions. While not every candidate has experience in medical devices specifically, any candidate can flourish in the medical device industry with the right set of qualifications and abilities. By looking for the traits that promise success… Read more »
The Medical Device Industry & Affordable Care Act – How Will It Impact Staffing?
The Affordable Care Act (ACA) has changed the healthcare landscape, but the long-term impact on the healthcare industry is still unknown. One often overlooked area of healthcare that is already feeling the effects of the ACA is the medical device industry. Job-market analysts are already predicting that the ACA will result in an increase in… Read more »


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Apr 21, 2012
to give the underwater feel to it.
I asked my assistant, Watson, how he like it, but he was too busy with his daily bath.
Posted by Barbara Strobel Lardon at 8:00 AM
Labels: Construction of Quilt, Creatures
4 comments:
- Terri Stegmiller said...
It looks great! I'm betting Watson would be much more interested if it was a real live fish.
- April 21, 2012 at 9:42 AM
- WoolenSails said...
Love the metallic for the thread, I can see glimmers of it here and there. I was looking at the thread in the store, but thought I should learn how to quilt with regular thread first;)
- April 21, 2012 at 12:20 PM
- Nellie's Needles said...
Looove it!
- April 21, 2012 at 1:26 PM
- Karen S said...
This is a beautiful quilt! And Watson is pretty cute too...
- April 21, 2012 at 4:12 PM
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Favorite Harry Potter Movie: Harry Potter and the Goblet of Fire...which is Year 4 at Hogwarts. It's also my favorite HP Book. Favorite String Instrument: oh Dang, my hillbilly is going to show with this answer , but it's a tie with a Banjo and a Fiddle/ViolinFavorite Country you want to visit: Japan...TokyoFavorite Breed of … Continue reading A Few Of My Favorite Things…
New Year, New Outing
We try to do something as a family when the New Year starts. I don't remember exactly when we started doing this...5 years ago? We tend to travel to St. Louis to do this family fun-fest because it offers a plethora of activities to choose from and it's only two hours south from the farm. … Continue reading New Year, New Outing
Up and Down The Chicago River
The water taxi from Chinatown to Michigan Avenue takes about 40 minutes. Chinatown in located on the near south side of Chicago and the river divides the city between north and south addresses. We were heading towards north Michigan Ave. The boat stops at a few different locations along the way, so you get to … Continue reading Up and Down The Chicago River
Planning A Road Trip To See A Wizard
I'm planning a trip for my family to go to Florida once school gets out in May. We want to go to see all of the Harry Potter sites that Universal Studios in Orlando has to offer. The kids LOVE the Harry Potter films (yes, we own all of the movies) and three out of … Continue reading Planning A Road Trip To See A Wizard
First Week Of 2018 Review
The first week of the new year, 2018, has already come and gone. Days are still cold and the nights even colder. In Illinois, we are used to the cold blowing across the prairie, causing the snow to drift into largish mountains of white. Blocking the view, hampering the walk to the mailbox. We haven't … Continue reading First Week Of 2018 Review


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As far as university creameries go, Utah State has one of the oldest. The original Utah State Creamery was located in the "Old Main" building from 1888 to 1918. At first, the creamery focused on butter and cheese, but in 1920 the university purchased equipment to manufacture ice cream. Aggie Ice Cream was born.
In 1921, Gustav Wilster came to Utah State's school and revolutionized the school's dairy department. His efforts resulted in students gaining a greater understanding of dairy production, manufacturing, and, much more importantly, how to make a higher quality ice cream.
Today, the creamery is located in Utah State's Nutrition and Food Sciences Building where visitors can enjoy over 25 flavors including Aggie Bull Tracks.
While Aggie Ice Cream is a favorite with students, a past Korean USU student appears to be the biggest fan. He acquired such an affection for Aggie Ice Cream during his time at the university that he devised a way to bring it back to his native country. With South Korean backers, he opened Aggie Korea Co., LTD. In 2004, the company was purchased by South Korea's electronics company, Electroland. Now the ice cream is known as "Munakee Shalle." Apparently, Aggie means "cute, healthy baby" in Korean. I guess everyone loves the look and smell of a cute, healthy baby, but the taste may be offputting.
Aggie Ice Cream also has the distinction of being the first ice cream to go in space. This was in large part thanks to USU's Aerospace Technologies Department. I did a bit of digging on this, but couldn't find any other information. I'm dying to know more facts. If you know anything like dates, flavors….ANYTHING, please let me know.


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Guiseley Architects
Guiseley Architects
Looking to find Architects in Guiseley McNeil Beechey O'Neill Architects ensure that you get Architects service in Guiseley and we would like to assist you, to develop those thoughts & ideas into a real possibility.
At McNeil Beechey O'Neill Architects LLP, we are able to provide our customers with a complete Architects service in Guiseley from preliminary appraisals to completion of works and even beyond. Throughout our 30+ years in practice as chartered architects covering Guiseley area, providing Architects Services we have built up a wealth of understanding assisting Guiseley homeowners, building contractors, project developers, businesses and organisations and also medical institutions, having to find Architects Services.
As a Chartered Practice of Architects, carrying out Architects Services in Guiseley you actually have the comfort of knowing we deliver our wide range of architectural design expertise consistent with the RIBA's Plan of Work, which you can refer to below. For many clients we provide a complete service from stages 1 to 6. Although we are very happy to discuss tailored Architects Services for a development in Guiseley, as we acknowledge the individualism of each client and their project, is a unique requirement that deserves our best attention.
McNeil Beechey O'Neill Architects LLP equally recognise that a construction project will often seem to be a complex undertaking, however, we do pride ourselves in becoming familiar with our clients' needs, guiding them through each stage of the –service—Services which we provide in Guiseley, and including design, planning and building processes.
We have a verified reputation of giving you a Architects Service, in Guiseley thoughtfully planned commercially realistic projects that satisfy the expectations of the users and our clients alike.
To put it succinctly our Architects expertise
Whatever your own project type, we offer a free of charge Guiseley based initial Architects Service Consultation. Whereby we determine {your|the client] requirements (the brief), explore constraints and also opportunities before supplying a fee quote that responds to the Architects products and services in Guiseley required by you the client. Naturally we will then help you through the next steps, whether that be conceptual drawings, planning drawings/consents, Building Regulations or getting your project on site in Guiseley.
Essentially our ability to give you a full Architects services in the Guiseley area enabling you to concentrate on something else. Knowing you can be confident mbo architects to deal with the steps in hand.
Mbo Architects cover the majority of areas of the UK, including Guiseley from our architects studio in York. For anybody who is a Householder, Contractor, House Builder, Design & Build Contractor, whether your project is Domestic, Residential, Hotel, Conservation and listed building, or Medical Practice to name a few. You can find a more samples of our works in our portfolio.
It doesn't matter if you are just after a bit of advice, on Architects Services in Guiseley area, feel free to call us, and we will be glad to guide you through the next steps.
First Steps in Guiseley – RIBA Stages 1+2
All working relationships start with a mutual exchange of views, regarding Architects Services in Guiseley. In the same way, the starting point in the Client Architect partnership is to get together in Guiseley to talk over how Architects Services will help your project. This will make it possible for us to establish a project brief outlining the range of the project together with the special requirements you might have. From the defined brief we can easily establish feasibility sketches for information. Your chosen sketch option can then be developed through regular discussion with you, to produce a series of images exploring the agreed proposals in plan form and 3D model/sketch.
Quite often a lot of these sketch proposals will also form part of the pre-application meetings with the Guiseley local planners to verify feasibility and consultation requirements in preparation for a full planning application.
Working with Planners – RIBA Stages 3+4
Mbo Architects will advance the sketch proposals into a comprehensive set of drawings in preparation for submitting and also gaining planning consent in Guiseley. During this stage we will conduct further discussions with the local Guiseleyplanning authority and other statutory bodies, as required in order to gain feedback on the proposals.
Once complete we can prepare and submit all the appropriate forms with the required drawings, as part of the Architects Service in Guiseley and papers, continue to manage the application process to obtain the necessary result.
Architects Service in Guiseley – Detail Design and Construction
RIBA Stages 5+6
As soon as the coveted planning approval has been gained we will further develop the drawings to a technical level, ready to submit for Building Regulations consent. For some household projects the Approved drawing at building regulation approval stage, could possibly be sufficient for a proficient building contractor to provide the required works. However, on more multifaceted projects and part of a full service we can easily further develop the drawings to create a comprehensive set of construction drawings with a specification. This level of detail can be sent out to a number of contractors to tender for the works.
For the period of construction we can administer the building contract on your behalf, undertaking regular site visits in Guiseley in order to check and ensure progress of the building works, consistent with the contract and Architects Service Provided by mbo architects.
If you need architects services, in Guiseley we are able to help you. Give us a call for more information.


text:
Caetano 25 Apr 1974
by World Federation of Democratic Youth
Budapest, Hungary
Undated, 1976?
Type: Poster
Coverage in Africa: Southern Africa
Language: English
Checklist of world dictators and dates on which they were overthrown. In addition to Marcelo Caetano of Portugal who was overthrown by the "Carnation Revolution" in April 1974, the poster shows the end of the rule of Dimitris Ioannidis (a leading figure in the Greek military junta of 1967–1974), Marshal Lon Nol of the Khmer Republic (1970-1975), President of South Vietnam Nguyễn Văn Thiệu (1965-1975), and Francisco Franco of Spain (who died in November 1975). The checklist names other leaders who have not yet been deposed: Pac Jeung Hi (Park Chung-hee of South Korea?), Augusto Pinochet of Chile, Ian Smith of Southern Rhodesia, and John Vorster of South Africa. [Budapest is the headquarters of the World Federation of Democratic Youth.] Size: 26-1/2 x 18-1/2 inches.
Collection: Private collection of David Wiley and Christine Root


text:
1: Introduction
2: Location, Borders, and Lakes
3: Geologic Structure and Landforms
4: Climate
5: Vegetation
6: Pre-historic and Early Historic Settlements
7: Survey Systems
8: Southern Hamlets, Villages, and Towns
9: Mennonite and Hutterite Settlements
10: First Nations Settlements
11: Northern Settlements
12: The Southern Cities
13: Mining and Oil Extraction
14: Agriculture
15: Industry / Manufacturing
16: Water Resources
17: Parks, Recreation, Sports
18: Transport and Communications: Past and Present
19: Legal Issues and Law EnforcementGlossary
Chapter 3: Geologic Structure and Landforms
Geologic interpretation of images begins with the identification of lineaments, "regional linear features caused by linear arrangement of regional morphological features such as streams, escarpments, and mountain ranges and tonal features that in many areas are the surface expressions of fractures or fault zones."[i] The emphasis in this section is on geologic structures and landforms, both of which can be easily identified on air photographs and other images.
The sequence followed here is that often found in geomorphology books. Geologic structures and structurally controlled landforms are illustrated first, followed by images of mass wasting and of the results of the agents of erosion—running water, ground water, ice, wind and the sea. The meandering rivers of southern
3.13: Slumping: Souris Gorge Between Souris and Wawanesa
East of Souris the southeastward flowing
The valley sides and the hummocky land 14 of the Tiger Hills are covered by deciduous woodland except for some areas cleared for cattle grazing 15. This contrasts with the flat land south 16 and north 17 of the valley that has been cleared for agriculture. These areas are underlain by glacial deposits, and to the north very minor northeast/southwest trending ridges 18 are picked out by the rain splash effect: rain splashes small-sized, dark-coloured fragments from crests, leaving behind coarse light-toned materials. To the west water-filled hollows 19 emphasize this topographic pattern.
Figure 3.13: Slumping: Souris Gorge Between Souris and Wawanesa
Figure 3.13
Vertical air photograph: A11046-57
Flight height: 9,520 feet a.s.l.; lens focal length: 6 inches
Date: May 21, 1947
Scale: 1:15,800 (approx.)
Location: Township 6, Range 18 WI
Map sheets: 1:250,000 62G Brandon
1:50,000 62G/12 Wawanesa


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The M-band is the transverse structure that cross-links the thick filaments in the center and provides a perfect alignment of the A-band in the activated sarcomere. The molecular composition of the M-bands in adult mouse skeletal muscle is fiber-type dependent. All M-bands in fast fibers contain M-protein while M-bands in slow fibers contain a significant proportion of the EH-myomesin isoform, previously detected only in embryonic heart muscle. This fiber-type specificity develops during the first postnatal weeks. However, the ratio between the amounts of myosin and of myomesin, taken as sum of both isoforms, remains nearly constant in all studied muscles. Ultrastructural analysis demonstrates that some of the soleus fibers show a diffuse appearance of the M-band, resembling the situation in the embryonic heart. A model is proposed to explain the functional consequence of differential M-band composition for the physiological and morphological properties of sarcomeres in different muscle types.
To compare training programs for women with trapezius myalgia regarding physical performance and pain, 102 women were randomized to strength, endurance, co-ordination and non-training groups. Before and after the intervention, static strength and dynamic muscular endurance in shoulder muscles were measured on a Cybex II dynamometer. Muscle activity in shoulder muscles was monitored via surface EMG. The signal amplitude ratio between the active and passive phase of repeated contractions indicated the ability to relax. Pain at present, pain in general and pain at worst were measured on visual analogue scales. After training, within group comparisons showed that the training groups rated less pain, and in the strength training group ratings of pain at worst differed from the non-training group. Using the non-training group as a reference, static strength increased in the strength and endurance training groups and muscular endurance in all training groups. The study indicates that regular exercises with strength, endurance or co-ordination training of neck/shoulder muscles might alleviate pain for women with work-related trapezius myalgia.
In 2005 the commonality of sarcotubular myopathy (STM) and limb girdle muscular dystrophy type 2H (LGMD2H) was demonstrated, as both are caused by the p D487N missense mutation in TRIM32 originally found in the Manitoba Hutterite population. Recently, three novel homozygous TRIM32 mutations have been described in LGMD patients. Here we describe a three generation Swedish family clinically presenting with limb girdle muscular weakness and histological features of a microvacuolar myopathy. The two index patients were compound heterozygotes for a frameshift mutation in TRIM32 (c.1560delC ) and a 30 kb intragenic deletion, encompassing parts of intron 1 and the entire exon 2 of TRIM32. In these patients, no full-length or truncated TRIM32 could be detected. Interestingly, heterozygous family members carrying only one mutation showed mild clinical symptoms and vacuolar changes in muscle. In our family, the phenotype encompasses additionally a mild demyelinating polyneuropathic syndrome. Thus STM and LGMD2H are the result of loss of function mutations that can be either deletions or missense mutations. (c) 2009 Wiley-Liss, Inc.
Patients with abnormal accumulations of desmin have been described in myopathies with or without cardiac involvement. Desmin deposits were sometimes associated with abnormal aggregates of other cytoskeletal proteins. In the present study we present how the cytoskeletal organisation of desmin, nestin, synemin, paranemin, plectin and alphaB-crystallin is altered in skeletal muscles from a patient with a L345P mutation in the desmin gene. In general, accumulations of desmin together with synemin, nestin, plectin and alphaB-crystallin were present between myofibrils and beneath the sarcolemma. However, as the biopsy samples were very myopathic, large variability in fibre size and fibre maturation was seen, thus the myofibrillar content and the cytoskeletal organisation varied considerably. In cultured satellite cells from the patient, desmin aggregates were not observed in initial passages, but occurred over time in culture in the form of perinuclear, peripheral or cytoplasmic deposits. Nestin colocalised to the abnormal desmin deposits to a larger extent than did vimentin. alphaB-Crystallin was only present in cells with a disrupted desmin network. Plectin was altered in a subset of cells with a disrupted desmin network, whereas synemin and paranemin were not detected. We conclude that the L345P desmin mutation has a profound influence on the cytoskeletal organisation both in vivo and in vitro, which reflects the pathogenesis of the desmin myopathy.
Mice lacking the gene encoding for the intermediate filament protein desmin have a surprisingly normal myofibrillar organization in skeletal muscle fibers, although myopathy develops in highly used muscles. In the present study we examined how synemin, paranemin, and plectin, three key cytoskeletal proteins related to desmin, are organized in normal and desmin knock-out (K/O) mice. We show that in wild-type mice, synemin, paranemin, and plectin were colocalized with desmin in Z-disc-associated striations and at the sarcolemma. All three proteins were also present at the myotendinous junctions and in the postsynaptic area of motor endplates. In the desmin K/O mice the distribution of plectin was unaffected, whereas synemin and paranemin were partly affected. The Z-disc-associated striations were in general no longer present in between the myofibrils. In contrast, at the myotendinous and neuromuscular junctions synemin and paranemin were still present. Our study shows that plectin differs from synemin and paranemin in its binding properties to the myofibrillar Z-discs and that the cytoskeleton in junctional areas is particularly complex in its organization.
Desmin, the main intermediate filament (IF) protein in skeletal and heart muscle cells, is of great importance as a part of the cytoskeleton. The IFs surround and interlink myofibrils, and connect the peripheral myofibrils with the sarcolemma. In myotendinous junctions and neuromuscular junctions of skeletal muscle fibres, desmin is enriched. In the heart, desmin is increased at intercalated discs, the attachment between cardiomyocytes, and it is the main component in Purkinje fibres of the conduction system. Desmin is the first muscle-specific protein to appear during myogenesis. Nevertheless, lack of desmin, as shown from experiments with desmin knockout (K/O) mice, does not influence myogenesis or myofibrillogenesis. However, the desmin knock-out mice postnatally develop a cardiomyopathy and a muscle dystrophy in highly used skeletal muscles. In other skeletal muscles the organization of myofibrils is remarkably unaffected. Thus, the main consequence of the lack of desmin is that the muscle fibres become more susceptible to damage. The loss of membrane integrity leads to a dystrophic process, with degeneration and fibrosis. In the heart cardiac failure develops, whereas in affected skeletal muscles regenerative attempts are seen. In humans, accumulations of desmin have been a hallmark for presumptive desmin myopathies. Recent investigations have shown that some families with such a myopathy have a defect in the gene coding for alphaB-crystallin, whereas others have mutations in the desmin gene. Typical features of these patients are cardiac affections and muscle weakness. Thus, mutations in the desmin gene is pathogenic for a distinct type of muscle disorder.
Myofibrillar remodelling with insertion of sarcomeres is a typical feature of biopsies taken from persons suffering of exercise-induced delayed onset muscle soreness. Here we studied the presence of the sarcomeric protein myotilin in eccentric exercise related lesions. Myotilin is a component of sarcomeric Z-discs and it binds several other Z-disc proteins, i.e. alpha-actinin, filamin C, F-actin and FATZ. Myotilin has previously been shown to be present in nemaline rods and central cores and to be mutated in limb girdle muscular dystrophy 1A (LGMD1A) and in a subset of myofibrillar myopathies, indicating an important role in Z-disc maintenance. Our findings on non-diseased muscle affected by eccentric exercise give new information on how myotilin is associated to myofibrillar components upon remodelling. We show that myotilin was present in increased amount in lesions related to Z-disc streaming and events leading to insertion of new sarcomeres in pre-existing myofibrils and can therefore be used as a marker for myofibrillar remodelling. Interestingly, myotilin is preferentially associated with F-actin rather than with the core Z-disc protein alpha-actinin during these events. This suggests that myotilin has a key role in the dynamic molecular events mediating myofibrillar assembly in normal and diseased skeletal muscle.
Obscurin is a giant protein (700-800 kDa) present in both skeletal muscles and myocardium. According to animal studies, obscurin interacts with myofibrillar Z-discs during early muscle development, but is translocalised to be predominantly associated with the M-bands in mature muscles. The proposed function for obscurin is in the assembly and organisation of myosin into regular A-bands during formation of new sarcomeres. In the present study, the precise localisation of obscurin in developing and mature normal human striated muscle is presented for the first time. We show that obscurin surrounded myofibrils at the M-band level in both developing and mature human skeletal and heart muscles, which is partly at variance with that observed in animals. At maturity, obscurin also formed links between the peripheral myofibrils and the sarcolemma, and was a distinct component of the neuromuscular junctions. Obscurin should therefore be regarded as an additional component of the extrasarcomeric cytoskeleton. To test this function of obscurin, biopsies from subjects with exercise-induced delayed onset muscle soreness (DOMS) were examined. In these subjects, myofibrillar alterations related to sarcomerogenesis are observed. Our immunohistochemical analysis revealed that obscurin was never lacking in myofibrillar alterations, but was either preserved at the M-band level or diffusely spread over the sarcomeres. As myosin was absent in such areas but later reincorporated in the newly formed sarcomeres, our results support that obscurin also might play an important role in the formation and maintenance of A-bands.
There is a paucity of data regarding the pathogenesis of chronic exertional compartment syndrome (CECS), its consequences for the muscles and the effects of treatment with fasciotomy. We analyzed biopsies from the tibialis anterior muscle, from nine patients, obtained during a decompressing fasciotomy and during follow-up 1 year later. Control biopsies were obtained from nine normal subjects. Muscle capillarity, fiber-type composition and fiber area were analyzed with enzyme- and immunohistochemistry and morphometry. At baseline, CECS patients had lower capillary density (273 vs 378 capillaries/mm(2), P=0.008), lower number of capillaries around muscle fibers (4.5 vs 5.7, P=0.004) and lower number of capillaries in relation to the muscle fiber area (1.1 vs 1.5, P=0.01) compared with normal controls. The fiber-type composition and fiber area did not differ, but focal signs of neuromuscular damage were observed in the CECS samples. At 1-year follow-up after fasciotomy, the fiber area and the number of fibers containing developmental myosin heavy chains were increased, but no enhancement of the capillary network was detected. Thus, morphologically, patients with CECS seemed to have reduced microcirculation capacity. Fasciotomy appeared to trigger a regenerative response in the muscle, however, without any increase in the capillary bed.
The morphological appearance of the vastus lateralis (VL) muscle from high-level power-lifters on long-term anabolic steroid supplementation (PAS) and power-lifters never taking anabolic steroids (P) was compared. The effects of long- and short-term supplementation were compared. Enzyme-immunohistochemical investigations were performed to assess muscle fiber type composition, fiber area, number of myonuclei per fiber, internal myonuclei, myonuclear domains and proportion of satellite cells. The PAS group had larger type I, IIA, IIAB and IIC fiber areas (p<0.05). The number of myonuclei/fiber and the proportion of central nuclei were significantly higher in the PAS group (p<0.05). Similar results were seen in the trapezius muscle (T) but additionally, in T the proportion of fibers expressing developmental myosin isoforms was higher in the PAS group compared to the P group. Further, in VL, the PAS group had significantly larger nuclear domains in fibers containing > or = 5 myonuclei. The results of AS on VL morphology in this study were similar to previously reported short-term effects of AS on VL. The initial effects from AS appear to be maintained for several years.
Power-lifters have hypertrophic muscle fibers with fissures seen in cross-sections, called as fiber splitting.Whether this phenomenon is due to real splitting or defective regeneration has not been settled. To elucidate this matter,we have examined biopsies from the trapezius and vastus lateralis of power lifters (P group) and power lifters self-administrating anabolic steroids (PAS group). For this purpose, immunohistochemical staining of serial cross -sections was used. The PAS group had significantly more fibers with fissures than the P group in the vastus lateralis (1.2%+/-0.95% vs 0.35+/-0.34, P < 0.05) but not in the trapezius muscle (1.7% in both groups). Serial sections revealed that the fibers with fissures changed their profile profoundly over short distances. Some such fibers had a mature staining profile, whereas other fibers indicated recent degeneration and/or regeneration. Activation of satellite cells and formation of aberrant segments were also evident. We conclude that the so-called split fibers are due to defect regeneration. Some fibers with fissures are the results of old events of segmental muscle fiber damage, whereas the others reflect an ongoing process. The normal regenerative process is most likely disturbed in power-lifters by their continuous training with repeated high mechanical stress on the muscles.
The aim of this study was to examine the effects of training on the structural characteristics of the trapezius muscle in women with work-related trapezius myalgia. Muscle biopsies were taken before and after 10 weeks of three different training programs (strength, endurance and coordination). Enzyme-immunohistochemical analysis was performed to assess muscle fibre types, fibre area, capillary supply and cytochrome c oxidase (COX) activity. There was an increase in the proportion of type IIA fibres in strength trained group (P < 0.05). Strength training elicited a preferential increase in the area of type II fibres (P < 0.05); both strength and endurance programs induced an increase in the number of capillaries around type I and IIA muscle fibres. Finally, all training programs induced a decrease in the proportion of COX-negative fibres. In conclusion, the trapezius muscle of women with neck and shoulder myalgia is characterised by a great potential of adaptation to physical exercise over a period of 10 weeks. The significant changes in the number of capillaries and the specific changes induced by training at the level of muscle fibres might well explain the improvement of muscle function.
A skeletal muscle fibre maintains its cytoplasmic volume by means of hundreds of myonuclei distributed along its entire length. Therefore it is hypothesised that changes in fibre size would involve modifications in myonuclear number. In this study, we have examined whether 10 weeks of strength training can induce changes in the number of myonuclei and satellite cells in female trapezius muscles. Biopsies were taken pre- and posttraining from the upper part of the descending trapezius muscle of nine subjects. Muscle samples were analysed for fibre area and myonuclear and satellite cell number using immunohistochemistry. There was a 36% increase in the cross-sectional area of muscle fibres. The hypertrophy of muscle fibres was accompanied by an approximately 70% increase in myonuclear number and a 46% increase in the number of satellite cells. Myonuclei number was positively correlated to satellite cell number indicating that a muscle with an increased concentration of myonuclei will contain a correspondingly higher number of satellite cells. The acquisition of additional myonuclei appears to be required to support the enlargement of multinucleated muscle cells following 10 weeks of strength training. Increased satellite cell content suggests that mitotic divisions of satellite cells produced daughter cells that became satellite cells.
The purpose of this study was to investigate the immunohistochemical expression of androgen receptors (AR) in human vastus lateralis and trapezius muscles and to determine whether long-term strength training and self-administration of androgenic-anabolic steroids are accompanied by changes in AR content. Biopsy samples were taken from eight high-level power-lifters (P), nine high-level power-lifters who used anabolic steroids (PAS) and six untrained subjects (U). Myonuclei and AR were visualised in cross-sections stained with the monoclonal antibody against AR and 4',6-diamidino-2-phenylindole. The proportion of AR-containing myonuclei per fibre cross-section was higher in the trapezius than in the vastus lateralis (P<0.05). In the trapezius, the proportion of AR-containing myonuclei was higher in P compared to U and in PAS compared to both P and U (P<0. 05). On the contrary, in the vastus lateralis, there were no differences in AR content between the three groups. Myonuclear number in both muscles was higher in P compared to U and in PAS compared to both P and U (P<0.05). In conclusion, AR content differs greatly between human neck and limb muscles. Moreover, the regulation of AR-containing myonuclei following training and self-administration of androgenic-anabolic steroids is muscle dependent.
PURPOSE: To investigate the composition of the fibers in human extraocular muscles (EOMs) with respect to the sarco(endo)plasmic reticulum Ca(2+)ATPases (SERCA)-1 and -2 and to investigate possible correlations between SERCA and myosin heavy chain (MyHC) composition. METHODS: EOM samples were processed for immunocytochemistry with monoclonal antibodies specific against SERCA1 (fast isoform), SERCA2 (slow isoform), or different MyHCs. A total of 1571 fibers were analyzed. Microsomal EOM fractions were analyzed with SDS-PAGE and immunoblots. RESULTS: The fast fibers, containing MyHCIIa, accounted for 79% of the fibers in the orbital layer (OL) and 74% in the global layer (GL). More than 99% of these fibers contained SERCA1, and 86% of them coexpressed SERCA1 and -2. Almost all slow fibers stained with SERCA2; 54% of those in the GL and all in the OL coexpressed SERCA1 and -2. Fifteen percent of the fibers in the GL and less than 1% in the OL were MyHCeom(pos)/MyHCIIa(neg) fibers. All these contained SERCA1 and in the OL also stained strongly with anti-SERCA2. Biochemically SERCA2 was more abundant than SERCA1. CONCLUSIONS: The human EOMs had a very complex pattern of expression of the major protein regulating fiber relaxation rate. The coexistence of SERCA1 and -2, together with complex mixtures of MyHCs in most of the fibers provide the human EOMs with a unique molecular portfolio that allows a highly specific fine-tuning regimen of contraction and relaxation.
PURPOSE: To investigate the myosin heavy chain (MyHC) composition of human extraocular (EOM) and levator palpebrae (LP) muscle fibers. METHODS: Adult human EOMs and LP were studied with SDS-PAGE, immunoblots, and immunocytochemistry, with antibodies against six MyHC isoforms. Myofibrillar adenosine triphosphatase (mATPase) and reduced nicotinamide adenine dinucleotide (NADH)-TR activity and fiber area were also determined. RESULTS: Most of the fibers in both layers stained strongly with anti-MyHCIIa. Approximately 14% of the fibers in the global layer and 16% in the orbital layer were labeled with anti-MyHCI. The remaining 24% of the fibers in the global layer and 3% in the orbital layer were not stained with either of these two antibodies, but were reactive to anti-MyHCeom (MyHCeom(pos)/MyHCIIa(neg) fibers). The fibers stained with anti-MyHCI had acid-stable mATPase activity, and the remainder of the fibers had alkaline-stable mATPase activity. Almost all the slow fibers stained with both anti-MyHCI and anti-MyHCslow tonic in both layers. Anti-MyHCalpha-cardiac stained approximately 26% of these slow fibers in the orbital layer and 7% in the global layer. Some slow fibers in both layers lacked staining with anti-MyHCslow tonic or with anti-MyHCalpha-cardiac. MyHCemb and/or MyHCeom were also present in some of the fibers of all the groups. The LP did not stain with anti-MyHCslow tonic. CONCLUSIONS: The present study revealed that the human EOMs have a very complex fiber type and MyHC composition and differ significantly from the EOMs of other species. The features of the LP were distinct from those of the four recti, the obliquus superior, and the limb muscles.
PURPOSE: To determine the laminin isoform composition of the basement membranes (BMs) in the human extraocular muscles (EOMs) and relate it to the fact that EOMs are spared in laminin alpha2-chain-deficient congenital muscular dystrophy. METHODS: Samples from adult human EOMs and limb muscle were processed for immunocytochemistry, with monoclonal antibodies against laminin chains (Ln) alpha1 to -5, beta1 and -2, and gamma1. Neuromuscular junctions (NMJs) were identified with acetylcholinesterase reaction. The capillary density was measured in sections stained with anti-Lnalpha5. RESULTS: The extrasynaptic BM of the EOM muscle fibers contained Lnalpha2, -beta1, -beta2, and -gamma1, and, in contrast to limb muscle, it also contained Lnalpha4 and -alpha5, to some extent. The distinct laminin composition of the EOMs was confirmed by the presence of Lutheran protein, an alpha5-chain-specific receptor not found in limb muscle. At the NMJs, there was increased expression of Lnalpha4 and expression of Lnalpha2, -alpha5, -beta1, -beta2, and -gamma1 was also maintained. The capillary density was very high (1050 +/- 190 capillaries/mm(2)) in the EOMs and significantly (P < 0.05) higher in the orbital (1170 +/- 180 capillaries/mm(2)) than in the global (930 +/- 110 capillaries/mm(2)) layer. CONCLUSIONS: The human EOMs showed important differences in laminin isoform composition and capillary density when compared with human limb muscle and muscles of other species. The presence of additional laminin isoforms other than laminin-2 in the BM of the extrasynaptic sarcolemma could partly explain the sparing of the EOMs in Lnalpha2-deficient congenital muscular dystrophy.
Human satellite cells (SCs) are heterogeneous with respect to markers for their identification in the niche between the muscle fibre plasma membrane and its basal lamina. We have previously shown that, in biopsies from highly competitive power lifters, power lifters with long-term use of anabolic steroids and a population of healthy sedentary men, antibodies against the neuronal cell adhesion molecule (NCAM) and the paired box transcription factor Pax7 together label 94% of the SCs, NCAM alone labels 4% and Pax7 alone labels 1%. In the present study, we have further studied these biopsies with four markers related to SC activation and differentiation. Our study unequivocally shows that staining for MyoD and myogenin are present in nuclei of SCs and of myoblasts and myotubes in areas of muscle fibre regeneration. Staining for c-Met was observed in a proportion of Pax7+ SCs. However, widespread labelling of the sarcolemma precluded the quantification of c-Met+/Pax7+ SCs and the use of c-Met as a reliable SC marker. Pax7+ SCs labelled by anti-Delta like1 (Dlk1) were present in all samples but in variable proportions, whereas muscle progenitor cells related to repair were Dlk1⁻. Staining for Dlk1 was also observed in Pax7⁻ interstitial cells and in the cytoplasm of some small muscle fibres. Interestingly, the proportion of Dlk1+/Pax7+ SCs was significantly different between the groups of power lifters. Thus, our study confirms that human SCs show marked heterogeneity and this is discussed in terms of SC activation, myonuclei turnover, muscle fibre growth and muscle fibre damage and repair.
Presently applied methods to identify and quantify human satellite cells (SCs) give discrepant results. We introduce a new immunofluorescence method that simultaneously monitors two SC markers (NCAM and Pax7), the basal lamina and nuclei. Biopsies from power-lifters, power-lifters using anabolic substances and untrained subjects were re-examined. Significantly different results from those with staining for NCAM and nuclei were observed. There were three subtypes of SCs; NCAM(+)/Pax7(+) (94%), NCAM(+)/Pax7(-) (4%) and NCAM(-)/Pax7(+) (1%) but large individual variability existed. The proportion of SCs per nuclei within the basal lamina of myofibres (SC/N) was similar for all groups reflecting a balance between the number of SCs and myonuclei to maintain homeostasis. We emphasise that it is important to quantify both SC/N and the number of SCs per fibre. Our multiple marker method is more reliable for SC identification and quantification and can be used to evaluate other markers of muscle progenitor cells.
The present study investigated potential age-related changes in human muscle spindles with respect to the intrafusal fiber-type content and myosin heavy chain (MyHC) composition in biceps brachii muscle. The total number of intrafusal fibers per spindle decreased significantly with aging, due to a significant reduction in the number of nuclear chain fibers. Nuclear chain fibers in old spindles were short and some showed novel expression of MyHC alpha-cardiac. The expression of MyHC alpha-cardiac in bag1 and bag2 fibers was greatly decreased in the A region. The expression of slow MyHC was increased in nuclear bag1 fibers and that of fetal MyHC decreased in bag2 fibers whereas the patterns of distribution of the remaining MyHC isoforms were generally not affected by aging. We conclude that aging appears to have an important impact on muscle spindle composition. These changes in muscle spindle phenotype may reflect an age-related deterioration in sensory and motor innervation and are likely to have an impact in motor control in the elderly.
Data on the myosin heavy chain (MyHC) composition of human muscle spindles are scarce in spite of the well-known correlation between MyHC composition and functional properties of skeletal muscle fibers. The MyHC composition of intrafusal fibers from 36 spindles of human biceps brachii muscle was studied in detail by immunocytochemistry with a large battery of antibodies. The MyHC content of isolated muscle spindles was assessed with SDS-PAGE and immunoblots. Four major MyHC isoforms (MyHCI, IIa, embryonic, and intrafusal) were detected with SDS-PAGE. Immunocytochemistry revealed very complex staining patterns for each intrafusal fiber type. The bag(1) fibers contained slow tonic MyHC along their entire fiber length and MyHCI, alpha-cardiac, embryonic, and fetal isoforms along a variable part of their length. The bag(2) fibers contained MyHC slow tonic, I, alpha-cardiac, embryonic, and fetal isoforms with regional variations. Chain fibers contained MyHCIIa, embryonic, and fetal isoforms throughout the fiber, and MyHCIIx at least in the juxtaequatorial region. Virtually each muscle spindle had a different allotment of numbers of bag(1), bag(2) and chain fibers. Taken together, the complexity in intrafusal fiber content and MyHC composition observed indicate that each muscle spindle in the human biceps has a unique identity.
The sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) is a membrane protein that plays a crucial role in muscle relaxation by transporting cytosolic Ca2+ into the lumen of the sarco/endoplasmic reticulum. In this study, the presence of SERCA1 and SERCA2 was investigated in human intrafusal fibers by immunocytochemistry. Nuclear bag1 fibers contained both SERCA1 and SERCA2 isoforms, with predominant staining seen with SERCA2 in the A and B regions. Most nuclear bag2 fibers also contained SERCA1 and SERCA2 isoforms and their coexistence frequently occurred in the A region. SERCA1 was present whereas SERCA2 was generally absent in the nuclear chain fibers. The staining intensity seen with the SERCA1 monoclonal antibody varied in the order of chain>bag1>bag2. The expression of SERCA1 isoform was found to correlate with the presence of fast myosin heavy chain (MyHC) isoform in nuclear chain fibers, whereas for nuclear bag fibers there was no such apparent correlation between patterns of expression of SERCA and MyHC isoforms. The phenotype revealed for the human bag fibers was very sophisticated and adapted to attain a very wide range of contraction and relaxation velocities.
Muscle spindle density is extremely high in the deep muscles of the human neck. However, there is a paucity of information regarding the morphology and immunoreactivity of these muscle spindles. The objective of this study was to investigate the intrafusal fiber content and to assess the myosin heavy chain (MyHC) composition of muscle spindles from human deep neck muscles. In addition to the conventional spindles containing bag(1), bag(2), and chain fibers (b(1)b(2)c spindle), we observed a number of spindles lacking bag(1) (b(2)c spindle) or bag(2) (b(1)c spindle) fibers. Both bag(1) and bag(2) fibers contained slow tonic MyHCs along their entire fiber length and MyHCI, MyHCIIa, embryonic, and alpha-cardiac MyHC isoforms along a variable length of the fibers. Fetal MyHC was present in bag(2) fibers but not in bag(1) fibers. Nuclear chain fibers contained MyHCIIa, embryonic, and fetal isoforms with regional variations. We also compared the present data with our previous results obtained from muscle spindles in human biceps brachii and the first lumbrical muscles. The allotment of numbers of intrafusal fibers and the MyHC composition showed some muscle-related differences, suggesting functional specialization in the control of movement among different human muscles.
The myosin heavy chain (MyHC) content in different parts of, two jaw opening muscle, the human lateral pterygoid and the digastric muscles of five young adult and five elderly subjects (mean age 22 and 73 years, respectively) was determined, using gel electrophoresis and immunohistochemical methods. The lateral pterygoid of both young and elderly contained predominantly slow MyHC, and fast A MyHC was the major fast isoform. In contrast, the digastric was composed of slow, fast A and fast X MyHCs in about equal proportions in both age groups. About half of the lateral pterygoid fibres contained mixtures of slow and fast MyHCs, often together with alpha-cardiac MyHC. In the digastric, co-existence of slow and fast MyHCs was rare, and alpha-cardiac MyHC was lacking. On the other hand, co-expression of fast A and fast X MyHCs was found more often in the digastric than in the lateral pterygoid. In both age groups about half of the digastric IIB fibres contained solely fast X MyHC. In the lateral pterygoid, type IIB fibres with pure fast X MyHC was found in only one subject. The lateral pterygoid in elderly showed a significant amount of fibres with solely fast A MyHC, which were occasionally found in young adults. In the digastric, no significant differences were found between young and elderly, although the muscles of elderly contained lower mean value of slow MyHC, as compared to that of young muscles. It is concluded that the lateral pterygoid and the digastric muscles differ not only in the MyHC composition but also in modifications of the MyHC phenotypes during aging, suggesting that they have separate roles in jaw opening function.
The morphology of the abductor pollicis brevis muscle was studied in 20 patients suffering from hand-arm vibration syndrome. The main morphological changes observed were centrally located myonuclei and fibre type grouping (found in all 20 muscle biopsies), angulated muscle fibres (found in 19 biopsies), ring fibres and regenerating fibres (found in 18 biopsies) and fibrosis (found in 17 biopsies). The observed abnormalities are believed to reflect damage to both the muscle fibres and the motor nerve. The changes were related to different vibration exposure parameters. The number of fibres demonstrating centrally located nuclei correlated significantly with the cumulative vibration exposure, while the number of angulated fibres correlated significantly with the total vibration exposure time. This indicates that the vibrating tools may cause direct damage to muscle fibres as well as nerves.
Hereditary myopathy with lactic acidosis (HML) is caused by an intron mutation in the iron-sulphur cluster assembly gene (ISCU) leading to incorporation of intron sequence into the mRNA. This results in a deficiency of Fe-S cluster proteins, affecting the TCA cycle and the respiratory chain. The proteins involved in the Fe-S machinery are evolutionary conserved and shown to be fundamental in all organisms examined. ISCU is expressed at high levels in numerous tissues in mammals, including high metabolic tissues like the heart, suggesting that a drastic mutation in the ISCU gene would be damaging to all energy-demanding organs. In spite of this, the symptoms in patients with HML are restricted to skeletal muscle, and it has been proposed that splicing events may contribute to the muscle specificity. In this study we confirm that a striking difference in the splicing pattern of mutant ISCU exists between different tissues. The highest level of incorrectly spliced ISCU mRNA was found in skeletal muscle, while the normal splice form predominated in patient heart. The splicing differences were also reflected at a functional level, where loss of Fe-S cluster carrying enzymes and accumulation of iron were present in muscle, but absent in other tissues. We also show that complete loss of ISCU in mice results in early embryonic death. The mice data confirm a fundamental role for ISCU in mammals and further support tissue-specific splicing as the major mechanism limiting the phenotype to skeletal muscle in HML.
We describe the mapping and identification of the gene for hereditary myopathy with lactic acidosis (HML). HML is characterized by low physical performance, resulting in physical exertion that causes early exhaustion, dyspnoea and palpitations. Using an autosomal recessive mode of inheritance, we mapped the trait to chromosome 12q23.3-24.11, with a maximum lod score of 5.26. The 1.6-Mb disease-critical region contained one obvious candidate gene-ISCU-specifying a protein involved in iron-sulphur cluster assembly. IscU is produced in two isoforms; one cytosolic and one mitochondrial, coded for by different splice variants of the ISCU gene. Mutational analysis of all exon and intron sequences as well as 1000 bp of the promoter of the ISCU gene revealed one intron mutation that was specific for the disease haplotype. The mutation is located in a region with homology to the interferon-stimulated response element (ISRE), but we could not see any effect of the mutation on expression levels in vitro or in vivo. We did, however, observe a drastic difference in the splicing pattern between patients and controls. In controls the mRNA was, as expected, mainly in the mitochondrial form, while in the patients a larger mRNA transcript was predominant. Sequencing of the product revealed that the mutation activates cryptic splice sites in intron 5 resulting in aberrant mRNA containing 100 bp of the intron. To conclude, our data strongly suggest that an intron mutation in the ISCU gene, leading to incorrectly spliced mRNA, is the cause of myopathy with lactic acidosis in this family.
Basement membranes (BMs) are an important part of the physiological microenvironment of pancreatic islet cells. In mouse islets, beta-cells interact directly with BMs of capillary endothelial cells. We have shown that in the human islets, the capillaries are surrounded by a double BM both in foetal and adult tissues. The endocrine islet cells are facing a BM that is separate from the endothelia. Laminins are the functionally most important component of BMs. The only laminin isoform present in the human endocrine islet BM is laminin-511 (previously known as laminin 10). The islet cells facing this BM have a strong and polarized expression of Lutheran glycoprotein, which is a well-known receptor for the laminin alpha 5 chain. Dispersed human islet cells adhere to purified human laminin-511 and the binding is equally effectively blocked by a soluble form of Lutheran as by antibody against integrin beta1. Our results reveal unique features of the BM structure of human islets, different from rodents. This information has potentially important implications for the generation of an optimal microenvironment for beta-cell function, proliferation and differentiation.
PURPOSE: To study the myosin heavy chain composition of the human extraocular muscles (EOMs) during development.
METHODS: EOMs from human fetuses of 8 to 22 weeks of gestation were studied with immunocytochemistry and gel electrophoresis. Antibodies specific against nine isoforms of myosin heavy chain (MyHC) were used in serial frozen sections.
RESULTS: The developing EOMs had a delayed time course of myotube formation and a unique composition and distribution of MyHCs compared with human limb skeletal muscle. The primary myotubes coexpressed two developmental isoforms of MyHCI from the earliest stages. The third developmental MyHCI delineated the future orbital layer at 10 to 12 weeks of gestation. MyHC-slow tonic also appeared early, whereas MyHC alpha-cardiac and MyHC-extraocular, important components of adult EOM, were never detected at the gestational ages studied.
CONCLUSIONS: The developmental features of the EOMs differed significantly from those reported for limb muscles of the corresponding ages. It is clear that the knowledge of limb muscle development does not fully apply to more specialized muscles, such as the eye muscles. The extreme complexity displayed by the EOMs probably reflects their distinct embryonic origin, innervation, and regulatory program of myogenesis.
In addition to being the specialized site for transmission of force from the muscle to the tendon, the myotendinous junction (MTJ) also plays an important role in muscle splitting during morphogenesis. An early event in the formation of the MTJ is a regional deposition of basement membranes. We used immunocytochemistry to investigate the distribution of laminin chains during the development of MTJs in human limb muscle at 8-22 weeks of gestation (wg) and in adult MTJs. We used polyclonal antibodies and a new monoclonal antibody (MAb) against the human laminin alpha1 G4/G5 domains. At 8-10 wg, laminin alpha1 and laminin alpha5 chains were specifically localized to the MTJ. Laminin alpha1 chain remained restricted to the MTJ at 22 wg as the laminin beta2 chain had appeared, whereas the laminin alpha5 chain became deposited along the entire length of the myotubes from 12 wg. In the adult MTJ, only vestigial amounts of laminin alpha1 and laminin alpha5 chains could be detected. On the basis of co-distribution data, we speculate that laminin alpha1 chain in the forming MTJ undergoes an isoform switch from laminin 1 to laminin 3. Our data indicate a potentially important role for laminin alpha1 chain in skeletal muscle formation.
Recent studies suggest important functions for laminin-8 (Ln-8; alpha4beta1gamma1) in vascular and blood cell biology, but its distribution in human tissues has remained elusive. We have raised a monoclonal antibody (MAb) FC10, and by enzyme-linked immunoassay (EIA) and Western blotting techniques we show that it recognizes the human Ln alpha4-chain. Immunoreactivity for the Ln alpha4-chain was localized in tissues of mesodermal origin, such as basement membranes (BMs) of endothelia, adipocytes, and skeletal, smooth, and cardiac muscle cells. In addition, the Ln alpha4-chain was found in regions of some epithelial BMs, including epidermis, salivary glands, pancreas, esophageal and gastric glands, intestinal crypts, and some renal medullary tubules. Developmental differences in the distribution of Ln alpha4-chain were detected in skeletal muscle, walls of vessels, and intestinal crypts. Ln alpha4- and Ln alpha2-chains co-localized in BMs of fetal skeletal muscle cells and in some epithelial BMs, e.g., in gastric glands and acini of pancreas. Cultured human pulmonary artery endothelial (HPAE) cells produced Ln alpha4-chain as M(r) 180,000 and 200,000 doublet and rapidly deposited it to the growth substratum. In cell-free extracellular matrices of human kidney and lung, Ln alpha4-chain was found as M(r) 180,000 protein.
Morphological properties of skeletal muscle were compared between wrist flexors and extensors within the same children (n = 8, six females, two males; age range 4 to 9y, median age 7 y) with wrist muscle imbalance secondary to spastic cerebral palsy (CP). Five patients had hemiplegic CP, two diplegic CP, and one patient had tetraplegic CP. Muscle biopsies were taken during either tendon transfer or tendon lengthening procedures. Analyses included distribution of muscle fibre types, fibre sizes, and expression of developmental myosins. Extensor fibre area was significantly greater than flexor fibre area for type 2A fibres and type 2B fibres but not for type 1 fibres. Coefficient of variation (CV) of fibre size for all three fibre types was greater for flexors compared with extensors. The greatest CV was observed for the type 2A fibres in flexors (39.5 [3.6%]). A wide variation was observed for expression of developmental myosin with the magnitude of the expression being greater, but not statistically significant, in flexors compared with extensors (5.4/mm2 vs 0.53/mm2). These data demonstrate that significant secondary myopathy of wrist flexor muscles results from CP.
Cultures of myoblasts isolated from cricopharyngeal muscles from patients with oculopharyngeal muscular dystrophy (OPMD) have been performed to study the effect of the expanded (GCG)8-13 repeat, located on the poly(A) binding protein nuclear-1 (PABPN1), on satellite cell phenotype. Cell cultures exhibited a reduced myogenicity, as well as a rapid decrease in proliferative lifespan, as compared to controls. The incorporation of BrdU decreased during the proliferative lifespan, due to a progressive accumulation of non-dividing cells. A lower fusion index was also observed, but myoblasts were able to form large myotubes when OPMD cultures were purified, although a rapid loss of myogenicity during successive passages was also observed. Myoblasts isolated from unaffected muscles did not show the defects observed in cricopharyngeal muscle cultures. The PABPN1 was predominantly located in nuclei of myoblasts and in both the nuclei and cytoplasm of myotubes in OPMD cultures. In vivo analysis of OPMD muscles showed that the number of satellite cells was slightly higher than that observed in age matched controls. Mutation of the PABPN1 in OPMD provokes premature senescence in dividing myoblasts, that may be due to intranuclear toxic aggregates. These results suggest that myoblast autografts, isolated from unaffected muscles, and injected into the dystrophic pharyngeal muscles, may be a useful therapeutic strategy to restore muscular function. Its tolerance and feasibility has been preclinically demonstrated in the dog.
Measuring the DNA telomere length of skeletal muscle in experienced endurance runners may contribute to our understanding of the effects of chronic exposure to endurance exercise on skeletal muscle. This study compared the minimum terminal restriction fragment (TRF) length in the vastus lateralis muscle of 18 experienced endurance runners (mean age: 42 +/- 7 years) to those of 19 sedentary individuals (mean age: 39 +/- 10 years). The runners had covered almost 50,000 km in training and racing over 15 years. Minimum TRF lengths measured in the muscle of both groups were similar (P = 0.805) and within the normal range. Minimum TRF length in the runners, however, was inversely related to their years spent running (r = -0.63, P = 0.007) and hours spent training (r = -0.52, P = 0.035). Therefore, since exposure to endurance running may influence minimum TRF length, and by implication, the proliferative potential of the satellite cells, chronic endurance running may be seen as a stressor to skeletal muscle.
Normal satellite cell cultures, isolated from human skeletal muscle, have a limited proliferative capacity and inevitably reach replicative senescence. In this study, we have focused on the consequences of a single oxidative stress by hydrogen peroxide (H(2)O(2)) on both proliferative capacity and myogenic characteristics. Treatment with 1mM H(2)O(2) for 30 min causes a small decrease in the viability and lifespan while the number of cells which are able to proliferate, decreases dramatically. This premature arrest of the cells in a non-proliferative state was not due to spontaneous differentiation since there was no increase in the number of myogenin positive cells. This stress did not affect the myogenicity of the cells or their ability to differentiate and fuse to form multinucleated myotubes. In addition, the mitotic clock does not seem to be modified by oxidative stress treatment since the rate of telomere shortening was similar in H(2)O(2)-treated and control cells. This could be the consequence of the high level of oxygen consumption with an even higher level of ROS being produced in skeletal muscle than in other tissues which would be counteracted by an increase in the antioxidant defense system.
In this study, we have investigated the consequences of aging on the regenerative capacity of human skeletal muscle by evaluating two parameters: (i) variation in telomere length which was used to evaluate the in vivo turn-over and (ii) the proportion of satellite cells calculated as compared to the total number of nuclei in a muscle fibre. Two skeletal muscles which have different types of innervation were analysed: the biceps brachii, a limb muscle, and the masseter, a masticatory muscle. The biopsies were obtained from two groups: young adults (23 +/- 1.15 years old) and aged adults (74 +/- 4.25 years old). Our results showed that during adult life, minimum telomere lengths and mean telomere lengths remained stable in the two muscles. The mean number of myonuclei per fibre was lower in the biceps brachii than in the masseter but no significant change was observed in either muscle with increasing age. However, the number of satellite cells, expressed as a proportion of myonuclei, decreased with age in both muscles. Therefore, normal aging of skeletal muscle in vivo is reflected by the number of satellite cells available for regeneration, but not by the mean number of myonuclei per fibre or by telomere lengths. We conclude that a decrease in regenerative capacity with age may be partially explained by a reduced availability of satellite cells.
Previous studies on aged animal muscle suggest that excitation-contraction uncoupling and fibre transitions play a central role in sarcopenia, the progressive loss and functional decline of aging skeletal muscle fibres. A drastic reduction in the voltage-sensing alpha1S-subunit of the transverse-tubular dihydropyridine receptor is believed to be the underlying cause for a decreased transmission of the surface depolarization signal into Ca2+-mediated muscle contraction. Extending these studies to human muscle, we asked whether potential changes in the relative expression of the voltage sensor occur in senescent human fibres. For internal standardization and as markers of potential fast-to-slow transitions, the fast isoforms of the Ca2+-binding element calsequestrin and the myosin heavy chain were employed. Besides small inter-individual variations in expression levels, the microsomal immunoblot analysis of vastus lateralis autopsy specimens from male humans aged 18 to 82 years of age showed no major changes in the relative abundance of the alpha1S- and alpha2-dihydropyridine receptor, fast calsequestrin and the slow/fast myosin heavy chains. The oligomeric status of the alpha1S-dihydropyridine receptor was unaltered in aged fibres. Biochemical assays revealed no significant modifications in Ca2+-ATPase activity and a reduced Ca2+-binding capacity in aged human muscle preparations. Although impairments of other Ca2+-regulatory proteins and/or disturbed protein-protein interactions might be involved in the pathophysiological changes of sarcopenia, dihydropyridine receptor and calsequestrin expression seem to be preserved during the aging process of human skeletal muscle fibres. Hence, the supposition that excitation-contraction uncoupling is responsible for sarcopenia can not be transferred from animal models to senescent human muscle without modifications.
We studied muscle spindles in the first lumbrical muscle of adult humans using myofibrillar ATPase (mATPase) activity. We found that muscle spindles exhibited a marked variability with respect to the number, position, length and detailed histochemical features of nuclear bag1, nuclear bag2 and nuclear chain fibers. Regarding mATPase activity, the nuclear bag2 fibers displayed lower alkali-stable mATPase activity along their length and many nuclear bag1 fibers tended to have lower acid-stable activity in the outer B region, whereas nuclear chain fibers exhibited medium acid-stable mATPase activity at pH 4.6. Almost 10% of spindle fibers displayed atypical features, as they were either located only at one spindle pole or exhibited mixed characteristics at either pole. The number of intrafusal fibers per spindle varied between 8 and 24. Strikingly, only 2 pairs from 22 muscle spindles had identical allotments of their intrafusal fibers. Muscle spindles in the first human lumbrical muscle contained more intrafusal fibers (12.3 +/- 4 per spindle on average) and especially relatively more nuclear bag fibers compared to other human skeletal muscles. Since each spindle apparently represents a unique morphological and physiological entity, the observed variability in the number and characteristics of intrafusal fibers in the first human lumbrical muscle likely reflects a wide range of finely tuned muscle spindle responses.
The muscle fibre composition of three human intrinsic tongue muscles, the longitudinalis, verticalis and transversus, was investigated in four anterior to posterior regions of the tongue using morphological and enzyme- and immunohistochemical techniques. All three muscles typically contained type I, IIA and IM/IIC fibres. Type I fibres expressed slow myosin heavy chain (MyHC), type II fibres fast MyHC, mainly fast A MyHC, whereas type IM/IIC coexpressed slow and fast MyHCs. Type II fibres were in the majority (60%), but regional differences in proportion and diameter of fibre types were obvious. The anterior region of the tongue contained a predominance of relatively small type II fibres (71%), in contrast to the posterior region which instead showed a majority of larger type I and type IM/IIC fibres (66%). In general, the fibre diameter was larger in the posterior region. This muscle fibre composition of the tongue differs from those of limb, orofacial and masticatory muscles, probably reflecting genotypic as well as phenotypic functional specialization in oral function. The predominance of type II fibres and the regional differences in fibre composition, together with intricate muscle structure, suggest generally fast and flexible actions in positioning and shaping the tongue, during vital tasks such as mastication, swallowing, respiration and speech. Copyright 2003 S. Karger AG, Basel
BACKGROUND: The authors recently described a new autosomal dominant myopathy (OMIM 605637 inclusion body myopathy 3) associated with a missense mutation in the myosin heavy chain (MyHC) IIa gene (MyHC IIa, Human Gene Map [HGM] locus MYH2). Young patients showed minor changes in their muscle biopsies, although dystrophic alterations and rimmed vacuoles with 15- to 20-nm tubulofilaments identical to those in sporadic inclusion body myositis (s-IBM) were observed in some of the adult (especially older) patients. The current study was undertaken to investigate the relation between expression of the mutant MyHC IIa and pathologic changes in muscle.
METHODS: The expression of MyHC IIa in nine muscle specimens from six individuals carrying the mutation was analyzed by immunohistochemistry, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and a new reverse transcriptase--PCR method to measure the relative abundance of the various MyHC transcripts.
RESULTS: Young patients with muscle weakness and minor pathologic changes in muscle expressed MyHC IIa at undetectable levels. MyHC IIa was expressed at high levels in adults with a progressive clinical course and dystrophic muscle changes. In these cases, a large number of muscle fibers were hybrids with expression of more than one MyHC isoform. Both MyHC IIa alleles were equally expressed. The relative level of MyHC IIa transcripts exceeded that of the corresponding protein, indicating an increased turnover of mutated protein. MyHC IIa expression was a consistent finding in muscle fibers with rimmed vacuoles.
CONCLUSIONS: The clear correlation between pathologic changes and expression of MyHC IIa indicates that defects in MyHC may lead not only to muscle weakness but also to muscle degeneration. The consistent expression of MyHC IIa in muscle fibers with rimmed vacuoles indicates that the breakdown of sarcomeric proteins is a key element in the pathogenesis of rimmed vacuoles of s-IBM type.
Myosin constitutes the major part of the thick filaments in the contractile apparatus of striated muscle. MYH7 encodes the slow/beta-cardiac myosin heavy chain (MyHC), which is the main MyHC isoform in slow, oxidative, type 1 muscle fibers of skeletal muscle. It is also the major MyHC isoform of cardiac ventricles. Numerous missense mutations in the globular head of slow/beta-cardiac MyHC are associated with familial hypertrophic cardiomyopathy. We identified a missense mutation, Arg1845Trp, in the rod region of slow/beta-cardiac MyHC in patients with a skeletal myopathy from two different families. The myopathy was characterized by muscle weakness and wasting with onset in childhood and slow progression, but no overt cardiomyopathy. Slow, oxidative, type 1 muscle fibers showed large inclusions consisting of slow/beta-cardiac MyHC. The features were similar to a previously described entity: hyaline body myopathy. Our findings indicate that the mutated residue of slow/beta-cardiac MyHC is essential for the assembly of thick filaments in skeletal muscle. We propose the term myosin storage myopathy for this disease.
Human satellite cells are indispensable for maintenance of human muscle mass, but their implications in the pathogenesis of sarcopenia and sarcopenic obesity are still under debate. Nevertheless, satellite cell activation upon exercise seems unequivocally together with adequate nutrition to be the most effective countermeasure for sarcopenia and sarcopenic obesity.


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Paying to Play: Travel Agents Up 2019 Advertising Budgetsby Cheryl Rosen
Sometimes you have to spend money to make money. While most travel agents rely on word-of-mouth and local networking, a profitable year has many agents testing the waters of paid advertising.
Lainey Melnick, for example, says her ads in the local paper have helped position her as a travel expert against a formidable new competitor. And Gary Smith credits his paid Facebook ads with helping to grow his business by 37 percent in 2018.
For Melnick, the push came from Departure Lounge, a tony and trendy storefront agency in her Austin, Texas, backyard that gobbled up lots of attention from the local media. Then one day, she was approached by a salesperson for five small local newspapers, offering her a chance to write a column about travel in return for her buying a quarter-page ad in each publication. Total cost: $299 per paper, or $1,500 a month.
"My strategy has always been to not have any overhead, so I never paid for advertising before. But I felt I really had to put some money into my brand so I wouldn't lose business to this competition," she said. And so, she gave it a try.
Sharing the cost
She quickly learned to share the wealth, or rather the cost. Last month, she wrote about the Royal Caribbean Cruise Line Genie. Then she used her ad on the page to highlight a RCCL promotion, with co-op advertising from the cruise line.
While it's hard to quantify how much business came from the ads and how much would have come in anyway, "I sold seven cabins, and I do know that I am thought of locally as an expert," she said. "And the main Austin newspaper called to do a story on me, so it led to free advertising, as well."
At Dream Vacations Travel Perks, meanwhile, owner Gary Smith says his outlay for paid ads on Facebook definitely has brought a positive return on investment (ROI). Instead of asking his travel partners for cash, he asks for a free cabin, which he then leverages on Facebook through a "register to win" promotion. That gives him the email addresses of hundreds of potential customers.
Smith has been growing his business through partnerships with associations and corporations, building co-branded websites that offer travel services to their members. "I have 60 or 70 of those," he said, "and 99 percent don't ask for anything in return other than that we take good care of their people."
The programs, whose customers range from giant tech companies to small churches, have proven so successful that he has two employees dedicated to marketing, and budgets $100,000 for salaries and $50,000 for "actual ad purchases and events."
Face-to-face is still important
Still, though, Smith believes that nothing builds a relationship like talking to a customer face-to-face. "Just placing an ad in a newspaper makes you just one of a thousand, but I want to establish myself as an expert," he said. "When it comes to marketing, I do a little of everything. We still go to our local home show for three days, to networking groups and local events. I say, 'I have 15 seconds to tell you how I can make your travel experience better.' I do my elevator spiel and I always end with, 'But don't believe a word I just said, make me prove it to you. Give me a chance to earn your trust.'"
For 2019, he is looking at more targeted Facebook advertising for his agency, which is based in a small town in Oregon. "I can't imagine there is anyone in Oregon who has more experience than we do selling river cruises," he said. "So, over the next five months, we are going to be paying to spread the word that there is no better place to buy river cruises than from us."
Connecting over wine and music
Many agents, next year, will follow a tried-and-true partnership route to find new customers. At Barefoot Travel Agency in Frisco, Texas, for example, co-owners Wendi Cartwright and Debbie Stevens have been extending their marketing budget by partnering with local wineries on themed trips to Europe. For next year, they will expand that to include breweries as well.
"Beer and wine and food and travel go very well together," Cartwright said, "so we are doing two wine-and-beer cruises, one in April and one in August."
A good partner to work with is someone who "has a good base to begin with, a good following and a good camaraderie, and that does events already," she said. "Then we have a road map: We help you with the blurbs about the trip, but you need to put it on your Facebook page; create a specialized travel page; have x amount of events; put it in your communications; include our flyer in your shipments of wine; and display our poster in your store."
Meanwhile, Laurie Schuss, owner of Cruise with Laurie, will combine group marketing with paid advertising for the first time next year. She will be hosting two big groups in 2019, a barbershop music cruise where the group will perform (for which 175 people already have signed up) and a new golf concierge program.
"We just took a half page ad to promote that cruise," she said. "This will be our first time advertising, and we hope it will pan out and bring us some customers."


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fax view fax(208) 336-9154 Address PO Box 2110
P.O. Box 2110
Boise, Idaho 83701
United States
P.O. Box 2110
Boise, Idaho 83701
United States
|Areas of Practice
|1) Business Law, 2) Mediation and Arbitration and 3) Estate Planning and Probate
|Admitted Year
|Whitman College (B.A., 1970); University of Denver
|Most recent firm
|Hopkins Roden Crockett Hansen & Hoopes, PLLC
Alan Singer is a partner in Morgan Lewis's Securities Practice. Mr. Singer advises companies with regard to a diverse range of corporate and securities matters.
Andrew Hamilton is a partner in Morgan Lewis's Business and Finance Practice. Mr. Hamilton practices in the areas of mergers and acquisitions, private equity and venture capital, securities law, and general corporate counseling.


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Reveal and reach customers behind
the data
Custobar is the premier customer data and marketing automation platform that combines online and offline data into a single, actionable customer 360° view.
Bring your app to life with push messages or send emails and smses to all your customers based on their relationship and experience with you.
You did dream of a perfect CDP. We created it.


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City of Hermosa BeachHome Menu
How to Pay a Parking Citation / Ticket
A parking citation may be paid:
- Online*: https://dspayments.com/HERMOSABEACH
- Via automated phone system*: 1-866-353-7146
*There is a $1 processing fee per transaction for the above options. Foreign credit cards are not accepted online or via automated phone system, but may be used below for payments processed at the Finance Cashier Office at City Hall.
- By credit card: the City accepts Mastercard, Visa and American Express--a 2.75% processing fee will apply to all credit card transactions. Complete a City Credit Card Authorization Form and select one of the options below:
- Mail or deliver to: City of Hermosa Beach - Finance Cashier, 1315 Valley Drive, Room 101, Hermosa Beach, CA 90254
- Place in the drop box by the front door of City Hall
- E-mail to: email@example.com
- Fax to: (310) 937-5959
- In person: payments may be made at the Finance Cashier Office (City Hall – 1315 Valley Drive, Room 101) during regular business hours Mon-Thu 7 AM to 6 PM. We accept Mastercard, Visa, American Express, checks, or cash. Please be advised that we do not accept unrolled coins in payment of amounts greater than $5.00. After hours payments are accepted in the drop box by the front door of City Hall (credit card authorization forms or checks only--no cash, please).
Low Income Installment Payment Plan
- Qualifying low-income individuals may apply for an installment payment plan.
- Please review the City's Low Income Payment Plan FAQs for details.
- To apply, fill out an Low Income Payment Plan Application and mail with appropriate back up documentation to the address noted on the form.
Questions? Please contact the Finance Cashier Office at (310) 318-0223 or e-mail: email@example.com
Contesting a Parking Citation / Ticket
Administrative Review Request - Step 1
- If you would like to contest a parking citation/ticket, please fill out and return the Request for Administrative Review form (.pdf) with any supporting documentation to the Finance Cashier Office within 21 days of issuance of the citation (see below for submittal instructions).
- The Hermosa Beach Police Department's Administrative Services Coordinator will review the information submitted and make a determination to uphold or dismiss your citation, and the result will be mailed to the address provided.
Administrative Hearing Request – Step 2 (if needed)
- If you do not agree with the results of the administrative review, you may fill out and return a Request for Administrative Hearing form (.pdf) to the Finance Cashier Office within 21 days of the mailing date of the result of the administrative review.
- In addition to submitting the Request for Administrative Hearing, you must pay the full amount of the parking penalty or claim indigence for an Administrative Hearing to be scheduled.
- To claim indigence, complete the Request for Non-Payment of Fines Application and submit to Finance Cashier within five days of requesting an Administrative Hearing. Claims of indigence are reviewed and approved by the Community Services Division. For more information, please read the Indigent Policy.
- Administrative hearings will be scheduled by appointment on alternating Thursdays between 3-5:50 PM at City Hall (1315 Valley Drive) where you will have the opportunity to provide testimony and evidence on your parking citation in front of a Hearing Officer.
- The Hearing Officer's decision to uphold or dismiss your citation is final and results will be mailed to the address provided.
Administrative Review and Hearing Request Forms with supporting documentation may be submitted via:
- Mail or delivered to (including payment for an Administrative Hearing): City of Hermosa Beach, Finance Cashier, 1315 Valley Drive, Room 101, Hermosa Beach, CA 90254.
- Place in the drop box by the front door of City Hall.
- E-mail to: firstname.lastname@example.com
- Fax to: (310) 937-5959
Administrative Hearings for citations are scheduled on alternating Thursdays, between 3-5:50 PM at the Hermosa Beach City Hall and are conducted by appointment only. Unscheduled cases will not be heard under any circumstances.
Appeal to Superior Court - Step 3 (if needed)
Note: According to California Vehicle Code Section 40230(a), you may not skip the Administrative Review or Administrative Hearing and file immediately for a Superior Court ruling.
You may appeal the decision of the Administrative Hearing Examiner to the Superior Court. You must:
- Request an appeal within 30 calendar days of the date of the Notice of Results of Administrative Hearing by submitting the applicable fees at Los Angeles Superior Court, 111 N. Hill Street, Los Angeles, CA 90012.
- Serve a copy of the Notice of Appeal upon the City either in person or by first-class mail within ten (10) calendar days of the filing of an appeal.
For more information and complete instructions regarding appeals, please visit LACourt.org.
- For general questions and submittals, please contact the Finance Cashier Office at 310-318-0223 or e-mail: email@example.com.
- For parking violation questions, please e-mail Hermosa Beach Police Department parking enforcement supervisors Sean Tanabe or Carlos Burgos (please do not e-mail Request for Administrative Review or Hearing forms).
- For questions on the status of an existing Administrative Review after a minimum processing time of 21 days, please e-mail Hermosa Beach Police Department Administrative Services Coordinator Ennis Jackson or contact (310) 318-0209.
Towed/Impounded Vehicle Release Procedures
To gain release of a towed or impounded vehicle, contact the Police Records Office at (310) 318-0360 (monitored 24/7). The Police Department will advise if a vehicle may be picked up directly from the tow yard or if any applicable fees, past parking citation payments and/or a vehicle Release Form must be obtained.
To obtain a Release Form:
- During regular business hours (Mon-Thu 7AM to 6PM):
- Visit the Finance Cashier Office at 1315 Valley Drive, Room 101
- Pay outstanding balance
- Obtain receipt for payment
- Take receipt of payment to the Police Records Office at 540 Pier Avenue
- Obtain Release Form (registered owner only)
- After hours/weekends (Mon-Thu before 7 AM or after 6 PM and Fri-Sun):
- Visit the Police Records Office at 540 Pier Avenue
- Pay outstanding balance (cash/exact change only)
- Obtain Release Form (registered owner only)
Location for Towed Vehicles – please check with the Police Records Office (310) 318-0360 prior to visiting to ensure all requirements for release have been met. Applicable towing, storage and other fees may be charged by the towing company prior to vehicle release.
- Bruffy's Tow
11101 Hindry Avenue
Los Angeles, CA 90045
(310) 395-0084
Impounded Bicycles & Scooters
The Hermosa Beach Police Department will provide release once applicable fees have been paid at the Finance Cashier's Office at 1315 Valley Drive, Room 101 during normal business hours (Monday to Thursday, 7 a.m. to 6 p.m) or at the Police Records Office at 540 Pier Avenue (open 24/7--cash/exact change only).


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You are here
PrOwlin.com is a social media site that maintains the best two-way line of communication for the players in nightlife industry.
Website: http://PrOwlin.com
Founded: 2012
Employees: 2
Quick Pitch:
PrOwlin.com is a social media website that provides a community for the players in the nightlife industry including merchants and guests. The service provided for the guests entails the most effective delivery of the most pertinent information for the guests needs. The website benefits the consumers by providing them with one location to access information, deals and promotions while giving merchants a vehicle to directly target their consumer.